Blockade of the galactose-binding sites of ricin by its linkage to antibody. Specific cytotoxic effects of the conjugates.
AuthorsThorpe PE,Ross WC,Brown AN,Myers CD,Cumber AJ,Foxwell BM,Forrester JT
JournalEuropean journal of biochemistry
PubMed ID6608447
A method is described for preparing specific cytotoxic agents by linking intact ricin to antibodies in a manner that produces obstruction of the galactose-binding sites on the B chain of the toxin and so diminishes the capacity of the conjugate to bind non-specifically to cells. The conjugates were synthesised by ... More
Enhancement of the thrombolytic potency of plasminogen activators by conjugation with clot-specific monoclonal antibodies.
AuthorsDewerchin M, Collen D
JournalBioconjug Chem
PubMed ID1790167
Detection of one attomole of [Arg8]-vasopressin by novel noncompetitive enzyme immunoassay (hetero-two-site complex transfer enzyme immunoassay).
AuthorsHashida S, Tanaka K, Yamamoto N, Uno T, Yamaguchi K, Ishikawa E
JournalJ Biochem (Tokyo)
PubMed ID1778973
'One attomole of [Arg8]-vasopressin (AVP) was detected by a novel noncompetitive enzyme immunoassay (hetero-two-site complex transfer enzyme immunoassay). AVP was indirectly biotinylated using N-hydroxysuccinimidobiotin and trapped onto an anti-AVP IgG-coated polystyrene ball. After washing, biotinylated AVP was eluted from the polystyrene ball with HCl and was reacted with 2,4-dinitrophenyl-fluorescein disulfide-bovine ... More
Activation of methotrexate-alpha-alanine by carboxypeptidase A-monoclonal antibody conjugate.
AuthorsHaenseler E, Esswein A, Vitols KS, Montejano Y, Mueller BM, Reisfeld RA, Huennekens FM
JournalBiochemistry
PubMed ID1731945
'Carboxypeptidase A (CP-A) and monoclonal antibody KS1/4 directed against an antigen on human lung adenocarcinoma cells (UCLA-P3) were derivatized by treatment with succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate and N-succinimidyl 3-(2-pyridyldithio)propionate, respectively. The derivatized proteins were reacted to produce thioether-linked enzyme-antibody conjugates. Sequential HPLC size-exclusion and DEAE chromatography separated the conjugate preparation from unreacted ... More
Selective destruction of nerve growth factor receptor-bearing cells in vitro using a hybrid toxin composed of ricin A chain and a monoclonal antibody against the nerve growth factor receptor.
AuthorsDiStefano PS, Schweitzer JB, Taniuchi M, Johnson EM
JournalJ Cell Biol
PubMed ID2993314
'A hybrid toxin composed of ricin A chain and a monoclonal antibody directed against the rat nerve growth factor (NGF) receptor (192-IgG) was prepared using the heterobifunctional cross-linking agent N-succinimidyl-3-(2-pyridyldithio)-propionate and purified by affinity chromatography. Characterization studies showed that the hybrid, 192-s-s-A, displaced bound 125I-labeled 192-IgG from rat superior cervical ... More
Basic studies on heterobifunctional biotin-PEG conjugates with a 3-(4-pyridyldithio)propionyl marker on the second terminus.
AuthorsKaiser K, Marek M, Haselgrübler T, Schindler H, Gruber HJ
JournalBioconjug Chem
PubMed ID9258454
'Heterobifunctional poly(ethylene glycol) (PEG) derivatives with a biotin terminus have been synthesized and characterized with respect to avidin binding. Unambiguous measurement of biotinyl and pyridyldithiopropionyl end groups was established by selecting suitable assays and introducing necessary modifications. Functional studies on the binding of biotin-PEG conjugates to avidin tetramers revealed much ... More
The agonist binding site on the bovine bradykinin B2 receptor is adjacent to a sulfhydryl and is differentiated from the antagonist binding site by chemical cross-linking.
AuthorsHerzig MC, Leeb-Lundberg LM
JournalJ Biol Chem
PubMed ID7657637
'Chemical cross-linking was used to analyze the binding sites for the agonist bradykinin (BK) and the antagonists NPC17731 and HOE140 on the bovine B2 bradykinin receptor. [3H]BK and [3H]NPC17731 bound with high affinity to the same B2 receptor in bovine myometrial membranes as determined by the total number of specific ... More
Acidification of the cytosol inhibits endocytosis from coated pits.
AuthorsSandvig K, Olsnes S, Petersen OW, van Deurs B
JournalJ Cell Biol
PubMed ID2887575
'Acidification of the cytosol of a number of different cell lines strongly reduced the endocytic uptake of transferrin and epidermal growth factor. The number of transferrin binding sites at the cell surface was increased in acidified cells. Electron microscopic studies showed that the number of coated pits at the cell ... More
High-level conjugation of chelating agents onto immunoglobulins: use of an intermediary poly(L-lysine)-diethylenetriaminepentaacetic acid carrier.
AuthorsManabe Y, Longley C, Furmanski P
JournalBiochim Biophys Acta
PubMed ID3756213
'Diethylenetriaminepentaacetic acid (DTPA), a strong chelating agent, was covalently linked to murine monoclonal anti-HLA IgG1 antibody (H-1) with the use of poly(L-lysine) (Mr 14,000) as a multivalent, intermediary carrier, via thiol-disulfide exchange reaction. The conjugates contained up to 42.5 mol DTPA per mol antibody, and retained over 90% of their ... More
Comparative studies of the preparation of immunoliposomes with the use of two bifunctional coupling agents and investigation of in vitro immunoliposome-target cell binding by cytofluorometry and electron microscopy.
'The two coupling agents SPDP (N-succinimidyl-3-(2-pyridyldithio)propionate) and SATA (N-succinimidyl-S-acetylthioacetate) were compared in their efficiency and feasibility to couple monoclonal antibodies (Abs) via thioether linkage to liposomes functionalized by various lipophilic maleimide compounds like N-(3-maleimidopropionyl)-N2-palmitoyl-L-lysine methyl ester (MP-PL), N-(3-maleimidopropionyl)phosphatidylethanolamide (MP-PE), N6-(6-maleimidocaproyl)-N2-palmitoyl-L-lysine methyl ester (EMC-PL), and N-(6-maleimidocaproyl)phosphatidylethanolamine (EMC-PE). The composition of the ... More
ATP-dependent association of nuclear proteins with isolated rat liver nuclei.
AuthorsImamoto-Sonobe N, Yoneda Y, Iwamoto R, Sugawa H, Uchida T
JournalProc Natl Acad Sci U S A
PubMed ID3368451
'In vitro association of Xenopus nucleoplasmin and mammalian nonhistone chromosomal high mobility group 1 (HMG1) protein with nuclei isolated from rat liver was examined. Efficient association of nuclear proteins with isolated nuclei requires ATP, HCO3-, and Ca2+. Association occurred at 33 degrees C but not at 4 degrees C. ATP ... More
Preparation of monomeric Fab'-horseradish peroxidase conjugate using thiol groups in the hinge and its evaluation in enzyme immunoassay and immunohistochemical staining.
AuthorsIshikawa E, Yoshitake S, Imagawa M, Sumiyoshi A
JournalAnn N Y Acad Sci
PubMed ID6372612
'Horseradish peroxidase and Fab'' were conjugated by using thiol groups in the hinge of Fab''. Maleimide or pyridyl disulfide groups were introduced into peroxidase by treatment with N-succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate or N-succinimidyl 3-(2-pyridyldithio)propionate and were allowed to react with thiol groups in the hinge of Fab''. The conjugates were obtained in ... More
Bispecific F (ab')2 monomer prepared with anti-CD3 and anti-tumor monoclonal antibodies is most potent in induction of cytolysis of human T cells.
AuthorsNitta T, Yagita H, Azuma T, Sato K, Okumura K
JournalEur J Immunol
PubMed ID2528460
'Induction of cytolytic activity of peripheral blood mononuclear cells towards target cells was studied by preparing bispecific F(ab'')2 which was composed of two Fab'' fragments, one of which was derived from anti-CD3 monoclonal antibody and the other from anti-tumor monoclonal antibody. After reduction of the interchange disulfide bonds of these ... More
Sequence-specific scission of DNA by the chemical nuclease activity of 1,10-phenanthroline-copper(I) targeted by RNA.
AuthorsChen CB, Gorin MB, Sigman DS
JournalProc Natl Acad Sci U S A
PubMed ID7683427
'RNAs modified with the chemical nuclease 1,10-phenanthroline-copper(I) can achieve the sequence-specific scission of single- and double-stranded DNA targets. The RNAs are prepared in vitro by using 5-(3-aminoallyl)-UTP as the sole source of UTP and can be readily modified with 1,10-phenanthroline by using N-succinimidyl 3-(2-pyridyl-dithio)propionate (SPDP) to cross-link the ligand to ... More
Quantitative analysis of derivatized proteins prepared with pyridyl disulfide-containing cross-linkers by high-performance liquid chromatography.
AuthorsNa DH, Woo BH, Lee KC
JournalBioconjug Chem
PubMed ID10077481
'Determination of the introduced moieties into derivatized proteins is an essential step in the preparation and quality control of chemically defined immunoconjugates. For the derivatized proteins using pyridyl disulfide-containing cross-linkers such as N-succinimidyl 3-(2-pyridyldithio)propionate (SPDP) and 4-succinimidyloxycarbonyl-alpha-methyl-alpha-(2-pyridyldithio)tolu ene (SMPT), the derivatization degree (ratio of pyridyl disulfide moieties to protein) has ... More
Fabrication of self-assembled protein A monolayer and its application as an immunosensor.
AuthorsLee W, Oh BK, Min Bae Y, Paek SH, Hong Lee W, Choi JW
JournalBiosens Bioelectron
PubMed ID14611753
'The self-assembled layer of modified protein A was fabricated. In order to modify protein A, the surface group of protein A was substituted with thiol (-SH) functionality by using N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP) and dithiothreitol (DTT). The formation of a self-assembled protein A layer on a Au substrate and its increased binding ... More
Monoclonal antibodies and a heterobifunctional reagent: a novel approach to the vectorial labeling of selected membrane proteins.
AuthorsJou YH, Schepart BS, Bankert RB
JournalImmunol Commun
PubMed ID7152550
'Many applications exist and others are envisioned for the chemical coupling of macromolecules to membrane proteins on the surface of mammalian cells. The ability to use antibody as a means to label and subsequently to follow the distribution of cell surface proteins is reported here. A new procedure is outlined ... More
Comparison of four bifunctional reagents for coupling peptides to proteins and the effect of the three moieties on the immunogenicity of the conjugates.
AuthorsPeeters JM, Hazendonk TG, Beuvery EC, Tesser GI
JournalJ Immunol Methods
PubMed ID2499636
'Peptide-carrier conjugates are widely used to raise antipeptide antibodies. In a model system using angiotensin and tetanus toxoid as the peptide and the carrier protein respectively, four cross-linking reagents were employed to study their effect on the immunogenicity of the conjugates. Optimization of the conjugation method for these heterobifunctional reagents, ... More
High affinity C10-Oeq ester derivatives of ryanodine. Activator-selective agonists of the sarcoplasmic reticulum calcium release channel.
'The plant alkaloids ryanodine and dehydroryanodine are specific and potent modulators of the sarcoplasmic reticulum calcium release channel. In the present study, acidic, basic, and neutral side chains esters of these diterpene compounds were prepared and their pharmacologic activities were assessed. Binding affinities of the novel C10-Oeq ester derivatives for ... More
Suppression of the antibody response to a polymorphic peptide from the platelet alloantigen integrin beta3 with low molecular weight antigen arrays.
AuthorsWatson DC, Reim J, Dintzis HM
JournalJ Immunol
PubMed ID8786303
'The allelic human platelet alloantigens PIA1/PIA2 are determined by a 33 Leu-Pro substitution in the second disulfide loop of the integrin beta3 subunit of the fibrinogen receptor, alphaIIb beta3 (GPIIb-IIIa). Alloantibodies to PIA1 cause neonatal alloimmune thrombocytopenia. We studied the suppression of specific Ab production to a disulfide-looped peptide spanning ... More
Molecular mimicry between an immunodominant amino acid motif on the 47-kDa lipoprotein of Treponema pallidum (Tpp47) and multiple repeats of analogous sequences in fibronectin.
AuthorsBaughn RE, Jiang A, Abraham R, Ottmers V, Musher DM
JournalJ Immunol
PubMed ID8752922
'Molecular mimicry, resulting from structural similarities between self-determinants on host Ags and an organism''s antigenic determinants (epitopes), can incite autoimmune events in certain bacterial and viral diseases. In the course of comprehensively mapping the 47-kDa lipoprotein (Tpp47) of Treponema pallidum subsp. pallidum using an overlapping synthetic peptide strategy, we identified ... More
Improvement of MRI probes to allow efficient detection of gene expression.
'Recently, it has been demonstrated that magnetic resonance imaging (MRI) utilizing monocrystalline iron oxide nanoparticles (MIONs) targeted to an engineered transferrin receptor enables imaging of gene expression. However, the relatively high doses of iron oxides used indicated the need for improved MR imaging probes to monitor changes in gene expression ... More
Effects of brefeldin A on endocytosis, transcytosis and transport to the Golgi complex in polarized MDCK cells.
AuthorsPrydz K, Hansen SH, Sandvig K, van Deurs B
JournalJ Cell Biol
PubMed ID1400572
'We have studied the effects of brefeldin A (BFA) on endocytosis and intracellular traffic in polarized MDCK cells by using the galactose-binding protein toxin ricin as a membrane marker and HRP as a marker of fluid phase transport. We found that BFA treatment rapidly increased apical endocytosis of both ricin ... More
Immunofluorescence measurement in a flow cytometer using low-power helium-neon laser excitation.
AuthorsShapiro HM, Glazer AN, Christenson L, Williams JM, Strom TB
JournalCytometry
PubMed ID6363017
'Helium-neon lasers are economical and efficient light sources; their utility in flow cytometry to date has been limited by the lack of fluorescent probes that can be excited at 633 nm. Allophycocyanin (APC), a highly fluorescent phycobiliprotein, can be used as an antibody label and has spectral characteristics suitable for ... More
Preparation and characterization of bifunctional biopolymers for receptor-based liposomal immunosensing.
AuthorsYu YY, Van Wie BJ, Koch AR, Moffett DF, Davis WC
JournalBiotechnol Prog
PubMed ID9548785
'In this study, we prepared bifunctional biopolymers for development of a novel liposomal immunosensing element. These biopolymers were produced such that a rat monoclonal antibody fragment Fab'' was linked to a cardiac protein Troponin I (TnI) peptide by a cross-linking reagent, o-phenylenedimaleimide (o-PDM) or N-sucinimidyl 3-(2-pyridyldithio)propionate (SPDP). The biopolymer formation ... More
Extended length heterobifunctional coupling agents for protein conjugations.
AuthorsBieniarz C, Husain M, Barnes G, King CA, Welch CJ
JournalBioconjug Chem
PubMed ID8741995
'A series of extended length heterobifunctional coupling agents is described. The successive aminocaproic acid homologation of succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate, a known 9-atom long maleimide active ester linker, yielded 16-, 23-, and 30-atom long maleimide active ester homologues. The performance study of these coupling agents in automated microparticle enzyme immunoassays showed that, ... More
Protein thiolation and reversible protein-protein conjugation. N-Succinimidyl 3-(2-pyridyldithio)propionate, a new heterobifunctional reagent.
AuthorsCarlsson J, Drevin H, Axén R
JournalBiochem J
PubMed ID708370
'A heterobifunctional reagent, N-succinimidyl 3-(2-pyridyldithio)propionate, was synthesized. Its N-hydroxysuccinimide ester group reacts with amino groups and the 2-pyridyl disulphide structure reacts with aliphatic thiols. A new thiolation procedure for proteins is based on this reagent. The procedure involves two steps. First, 2-pyridyl disulphide structures are introduced into the protein by ... More
Sensitive non-radioactive dot-blot hybridization using DNA probes labelled with chelate group substituted psoralen and quantitative detection by europium ion fluorescence.
AuthorsOser A, Roth WK, Valet G
JournalNucleic Acids Res
PubMed ID3344204
'A new labelling method for cloned DNA probes used in hybridization assays is described. The DNA insert of recombinant plasmid DNA was made partially single-stranded for the labelling reaction by a restriction enzyme digest, followed by a controlled exonuclease III incubation. A thiol-containing psoralen derivative was covalently bound through irradiation ... More
Comparison of conjugation procedures for the preparation of monoclonal antibody-enzyme conjugates.
AuthorsJeanson A, Cloes JM, Bouchet M, Rentier B
JournalJ Immunol Methods
PubMed ID3135335
'Four monoclonal antibodies belonging to different subclasses and with differing isoelectric points were coupled to horseradish peroxidase (HRP) and alkaline phosphatase (AP) using various conjugation procedures. The conjugates were tested by enzyme immunoassay and their efficiency was characterized by the antibody and enzyme concentrations needed to obtain an arbitrary OD ... More
Selective translocation of the A chain of diphtheria toxin across the membrane of purified endosomes.
AuthorsBeaumelle B, Bensammar L, Bienvenüe A
JournalJ Biol Chem
PubMed ID1597480
'Translocation is a necessary and rate-limiting step for diphtheria toxin (DT) cytotoxicity. We have reconstituted DT translocation in a cell-free system using endosomes purified from lymphocytes and have demonstrated this using two different probe/cell systems, which provided identical results: 125I-DT/human CEM cells and 125I-transferrin-DT/mouse BW cells. The cell-free DT translocation ... More
Specific enhancement of drug delivery to AKR lymphoma by antibody-targeted small unilamellar vesicles.
AuthorsMatthay KK, Heath TD, Papahadjopoulos D
JournalCancer Res
PubMed ID6713389
'Antibody targeting of drug-containing liposomes to specific cell populations provides the opportunity to improve cancer chemotherapy. We report here the efficacy of targeted liposomes containing methotrexate-gamma-aspartate against two murine T-lymphomas, AKR/J SL2 and R1.1. Both large and small unilamellar vesicles conjugated to anti-Thy-1.1 antibody associated with AKR lymphoma cells in ... More
Liposome immunoassay (LIA) with antigen-coupled liposomes containing alkaline phosphatase.
AuthorsKim CK, Lim SJ
JournalJ Immunol Methods
PubMed ID8445242
'Immunoliposomes were prepared and their immunoassay applications investigated. Liposomes were prepared from cholesterol and phospholipids including maleimidobenzoylphosphatidylethanolamine (MBPE) for conjunction with thiol-containing antigens. Alkaline phosphatase (APase) was entrapped in the liposome and BSA, the antigen, was modified by reaction with 3-(2-pyridyl-dithio)propionyl N-hydroxysuccinimide ester (SPDP) to introduce thiol groups for efficient ... More
Surface-functionalized nanoparticle library yields probes for apoptotic cells.
AuthorsSchellenberger EA, Reynolds F, Weissleder R, Josephson L
JournalChembiochem
PubMed ID14997519
'We have developed techniques for the efficient synthesis and screening of small libraries of surface-functionalized nanoparticles for the recognition of specific types of cells. To illustrate this concept we describe the development of a nanoparticle that preferentially recognizes apoptotic Jurkat cells in a manner similar to the apoptosis-recognizing protein annexin ... More
A silicon sensor-based filtration immunoassay using biotin-mediated capture.
'A sensitive sandwich immunoassay for human chorionic gonadotropin (hCG) was developed with biotin-mediated filtration capture and silicon sensor detection. A high density of biotin on the membrane assured efficient capture of complexes containing streptavidin and analyte. Capture efficiency was not affected over a wide range of filtration flow rates or ... More
Three-color immunofluorescence analysis of mouse B-lymphocyte subpopulations.
AuthorsParks DR, Hardy RR, Herzenberg LA
JournalCytometry
PubMed ID6370630
'We have modified a fluorescence-activated cell sorter (FACS) to make three independent immunofluorescence measurements on each cell and used this system to study mouse B-lymphocyte subpopulations. An argon-ion laser (emitting at 488 nm) excites fluorescein- and phycoerythrin-labeled reagents, and a tunable dye laser charged with rhodamine 6G (emitting at 615 ... More
Gene transfer from targeted liposomes to specific lymphoid cells by electroporation.
AuthorsMachy P, Lewis F, McMillan L, Jonak ZL
JournalProc Natl Acad Sci U S A
PubMed ID3186704
'Large unilamellar liposomes, coated with protein A and encapsulating the gene that confers resistance to mycophenolic acid, were used as a model system to demonstrate gene transfer into specific lymphoid cells. Protein A, which selectively recognizes mouse IgG2a antibodies, was coupled to liposomes to target them specifically to defined cell ... More
Covalent binding of antibodies to liposomes using a novel lipid derivative.
AuthorsGoundalkar A, Ghose T, Mezei M
JournalJ Pharm Pharmacol
PubMed ID6146693
'N-[3-(2-Pyridyldithio) propionyl] stearylamine (PDP-SA) was synthesized from a reaction between stearylamine and the heterobifunctional reagent N-succinimidyl-3-(2-pyridyldithio) propionate. Use of this PDP-SA to covalently couple antibodies to liposomes was investigated. The binding efficiency was found to be 24-32%. The antibodies bound to liposomes were shown to retain the specific antibody activity. ... More
Antibody-targeted liposomes: increase in specific toxicity of methotrexate-gamma-aspartate.
AuthorsHeath TD, Montgomery JA, Piper JR, Papahadjopoulos D
JournalProc Natl Acad Sci U S A
PubMed ID6572395
'Liposomes conjugated with anti-H2Kk antibody associate with L929 murine fibroblasts in 6- to 20-fold greater amount than do nonspecific liposomes. The ability of methotrexate-gamma-aspartate to inhibit L929 growth is increased 10-fold when encapsulated in targeted liposomes but is decreased to 50% when encapsulated in liposomes with no specificity for the ... More
A brief survey of methods for preparing protein conjugates with dyes, haptens, and cross-linking reagents.
AuthorsBrinkley M
JournalBioconjug Chem
PubMed ID1616945
Enhanced fluorescence in indirect immunophenotyping by the use of fluorescent liposomes.
AuthorsGray AG, Morgan J, Linch DC, Huehns ER
JournalJ Immunol Methods
PubMed ID2502576
'Small unilamellar liposomes were optimised for cell phenotyping by indirect immunofluorescence. This involved selection and covalent attachment to the liposome of a polyspecific ligand for cell-bound antibody. For this purpose sheep anti-mouse antibody was preferred to protein A because of its ability to attach to cell-bound IgG1 as well as ... More
New colored and fluorescent amine substrates for activated fibrin stabilizing factor (Factor XIIIa) and for transglutaminase.
AuthorsLorand L, Parameswaran KN, Velasco PT, Hsu LK, Siefring GE
JournalAnal Biochem
PubMed ID6193738
'Two fluorescent (FITC and 6-chloro-2-methoxyacridine) and an intensely colored (dabsyl) derivative of cadaverine were synthesized, following earlier work from this laboratory with dansyl-cadaverine, in order to enlarge the scope of possibilities for labeling some gamma-glutamine sites in proteins. Enzyme affinities of the amine substrates for human Factor XIIIa and for ... More
Demonstration of B-cell maturation in X-linked immunodeficient mice by simultaneous three-colour immunofluorescence.
AuthorsHardy RR, Hayakawa K, Parks DR, Herzenberg LA
JournalNature
PubMed ID6358897
'CBA/N mice carrying the X-linked immune deficiency gene (xid) have fewer splenic B cells than normal CBA mice and are unresponsive to a certain class of antigens. Studies of B-cell surface-marker expression and immune responsiveness have led to the commonly accepted idea that the B cells in adult xid mice ... More
ATP-dependent translocation of ricin across the membrane of purified endosomes.
AuthorsBeaumelle B, Alami M, Hopkins CR
JournalJ Biol Chem
PubMed ID7901210
'Ricin translocation was demonstrated (using both fluorescence- and radiolabel-based assays) across the membrane of endosomes purified from mouse lymphocytes. Selectivity of the process was shown by the absence of translocation activity of transferrin and horseradish peroxidase used as membrane-bound and fluid-phase endosome labels, respectively. Endocytosed 125I-ricin translocation was found to ... More
Transcytosis of GCSF-transferrin across rat alveolar epithelial cell monolayers.
AuthorsWidera A, Kim KJ, Crandall ED, Shen WC
JournalPharm Res
PubMed ID12948021
'PURPOSE: The purpose of this study was to use primary cultured rat alveolar epithelial cell monolayers to examine the potential of using transferrin receptor (TfR)-mediated transcytosis for noninvasive systemic protein drug delivery via the pulmonary route. METHODS: Freshly isolated rat type II pneumocytes were plated onto tissue culture-treated polycarbonate 12-mm ... More
Sensitivity and selectivity of ricin toxin A chain-monoclonal antibody 791T/36 conjugates against human tumor cell lines.
'Ricin toxin A chain (RTA) was conjugated to monoclonal antibody 791T/36, which was raised originally against human osteogenic sarcoma cell line 791T. The resultant conjugates were characterized and tested for cytotoxicity against a panel of human tumor cell lines representing a defined range of antigenicity with regard to 791T/36. Conjugates ... More
Endocytosis from coated pits of Shiga toxin: a glycolipid-binding protein from Shigella dysenteriae 1.
AuthorsSandvig K, Olsnes S, Brown JE, Petersen OW, van Deurs B
JournalJ Cell Biol
PubMed ID2564398
'Evidence is presented that endocytosis is involved in the transport to the cytosol of the cytotoxin from Shigella dysenteriae 1, Shiga toxin, which acts by removal of a single adenine residue in 28-S ribosomal RNA. Inhibition of endocytosis by ATP depletion of the cells prevented toxin uptake. Exposure of HeLa ... More
Transcellular processing of disulfide- and thioether-linked peroxidase--polylysine conjugates in cultured MDCK epithelial cells.
AuthorsWan JS, Persiani S, Shen WC
JournalJ Cell Physiol
PubMed ID2211845
'Horseradish peroxidase (HRP) was conjugated to nondegradable polycationic poly(D-lysine) (PDL) through either a thioether (HRP-S-PDL) or a disulfide (HRP-SS-PDL) linkage. The binding and transcytosis of these conjugates was studied in Madin-Darby canine kidney (MDCK) cell monolayers grown on 3-microns microporous polycarbonate filters. Conjugation of HRP to PDL with both linkages ... More
Gene transfer into human hepatoma cells by receptor-associated protein/polylysine conjugates.
AuthorsKim TG, Kang SY, Kang JH, Cho MY, Kim JI, Kim SH, Kim JS
JournalBioconjug Chem
PubMed ID15025528
'Receptor-associated protein (RAP) is a ligand for all members of low-density lipoprotein (LDL) receptor families. RAP is internalized into cells via receptor-mediated endocytic trafficking, making it an attractive mechanism for efficient gene delivery. In this study, we have developed a gene delivery system using RAP as a targeting ligand. A ... More
Homogeneous liposome immunoassay for insulin using phospholipase C from Clostridium perfringens.
AuthorsLim SJ, Kim CK
JournalAnal Biochem
PubMed ID9126376
'A new homogeneous liposome immunoassay system was developed in which analyte-phospholipase C conjugates are used instead of complement or melittin. This system was applied for the determination of insulin. Liposomes incorporated with calcein as a marker were prepared by the reverse-phase evaporation method. The lysis of liposomes was determined by ... More
A spectrophotometric method for the estimation of amino groups on polymer supports.
AuthorsGaur RK, Gupta KC
JournalAnal Biochem
PubMed ID2817353
'A simple and sensitive method for the estimation of polymer-supported amino groups is reported. The polymer support is treated either with N-succinimidyl-4-O-(4,4''-dimethoxytrityl)-butyrate or 2,4-dinitrophenyl-4-O-(4,4''-dimethoxytrityl)-butyrate and a catalytic amount of 4-dimethylaminopyridine. After removal of the excess reagent through washing, a weighed quantity of the polymer support is treated with perchloric acid ... More
Effective avidin coating of erythrocytes by disulfide bond formation. Production of a stable rosetting agent for multiple applications.
AuthorsFürfang J, Thierfelder S
JournalJ Immunol Methods
PubMed ID3522744
'A method is described for covalently coupling avidin to the membrane of glutaraldehyde-treated erythrocytes. The method is based on the use of the heterobifunctional coupling reagent N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP) and appears to be more effective than other protein coupling techniques. The resulting avidin-coated red cells are very stable and can be ... More
Controlled binding of liposomes to cultured cells by means of lectins.
AuthorsLiautard JP, Vidal M, Philippot JR
JournalCell Biol Int Rep
PubMed ID3841030
'Lectin-mediated binding of liposomes to Hela cells was analyzed as a function of different parameters. We show that the amount of lectin covalently bound to liposomes can be accurately controlled. We chose to work with 500 - to 1 000 molecules of WGA bound per liposome of 1 micron diameter. ... More
Selective killing of T lymphocytes by phototoxic liposomes.
AuthorsYemul S, Berger C, Estabrook A, Suarez S, Edelson R, Bayley H
JournalProc Natl Acad Sci U S A
PubMed ID3491992
'Two-fold specificity in drug delivery obtained through the localized activation of drugs by physical means and the attachment of drugs to proteins that bind to target cells might be used for highly selective cancer chemotherapy or for immunosuppression. Toward this end, a monoclonal antibody against an antigen on the surface ... More
Chemical modification of proteins: comments and perspectives.
AuthorsFeeney RE
JournalInt J Pept Protein Res
PubMed ID3570660
'The use of chemical modification of proteins has increased exponentially during the past two decades. Today the many different uses of chemical modification include determination of relative reactivities of side chain groups, the quantitation of individual amino acids, development of affinity reagents, mechanism-based reagents for pharmaceutical uses, cross-linking reagents, special ... More
Specific lysis of human tumor cells by T cells coated with anti-T3 cross-linked to anti-tumor antibody.
AuthorsPerez P, Titus JA, Lotze MT, Cuttitta F, Longo DL, Groves ES, Rabin H, Durda PJ, Segal DM
JournalJ Immunol
PubMed ID2944946
'Heteroaggregates containing anti-T3 cross-linked to anti-target cell antibodies have been shown to cause human T cells to lyse target cells that express antigens recognized by the anti-target cell antibody. In this study, we test targeted human T cells for the ability to lyse human tumor cells as a first step ... More
Protein-liposome conjugates with defined size distributions.
AuthorsLoughrey HC, Wong KF, Choi LS, Cullis PR, Bally MB
JournalBiochim Biophys Acta
PubMed ID2207121
'Conjugation of protein to liposomes by two coupling protocols is shown to result in vesicle aggregation. The degree of aggregation is directly related to the levels of protein conjugated to the liposomes. In an attempt to develop a method of generating stable, homogeneously sized protein-conjugated vesicles, highly aggregated liposome-protein conjugates ... More
Cobra venom factor immunoconjugates: effects of carbohydrate-directed versus amino group-directed conjugation.
AuthorsZara J, Pomato N, McCabe RP, Bredehorst R, Vogel CW
JournalBioconjug Chem
PubMed ID7578355
'Human IgM monoclonal antibody 16-88, derived from patients immunized with autologous colon carcinoma cells, was derivatized with two different cross-linkers, S-(2-thiopyridyl)-L-cysteine hydrazide (TPCH), which is carbohydrate-directed, and N-succinimidyl-3-(2- pyridyldithio)propionate (SPDP), which is amino group-directed. Two antibody functions, antigen binding and complement activation, were assayed upon derivatization with TPCH and SPDP. ... More
Preparation of four daunomycin-monoclonal antibody 791T/36 conjugates with anti-tumour activity.
AuthorsGallego J, Price MR, Baldwin RW
JournalInt J Cancer
PubMed ID6376376
'As an approach to developing more specific anti-tumour therapeutic agents, daunomycin has been covalently linked to the human tumour localizing, murine monoclonal antibody 791T/36. Four procedures for coupling drug to antibody were investigated. The sugar amino group of daunomycin was modified by reaction with succinic anhydride or cis aconitic anhydride ... More
Methods for the attachment of haptens and proteins to erythrocytes.
Preparation and application of antibodies coupled to the A chain of ricin.
AuthorsKrolick KA, Uhr JW, Vitetta ES
JournalMethods Enzymol
PubMed ID6346013
Antibody-directed liposomes: the development of a cell-specific cytotoxic agent.
AuthorsHeath TD, Bragman KS, Matthay KK, Lopez-Straubinger NG, Papahadjopoulos D
JournalBiochem Soc Trans
PubMed ID6724119
Targeting of cytotoxic cells with heterocrosslinked antibodies.
AuthorsSegal DM, Wunderlich JR
JournalCancer Invest
PubMed ID3284611
Preparation of [3-(2-pyridyldithio)propionoyl]insulins and horseradish peroxidase-insulin conjugates by high-performance liquid chromatographic separation.
AuthorsZaitsu K, Nakayama M, Ohkura Y
JournalChem Pharm Bull (Tokyo)
PubMed ID3046762
A simple method for the introduction of thiol group at 5'-termini of oligodeoxynucleotides.
AuthorsGaur RK, Sharma P, Gupta KC
JournalNucleic Acids Res
PubMed ID2740239
Targeting of the antiviral protein from Phytolacca americana with an antibody.
AuthorsMasuho Y, Kishida K, Hara T
JournalBiochem Biophys Res Commun
PubMed ID7092866
Affinity targeting of membrane vesicles to cell surfaces.
AuthorsGodfrey W, Doe B, Wallace EF, Bredt B, Wofsy L
JournalExp Cell Res
PubMed ID7026265
Coupling of monoclonal antibodies with enzymes.
AuthorsHaugland RP
JournalMethods Mol Biol
PubMed ID7550686
Preparation of avidin conjugates.
AuthorsHaugland RP, Bhalgat MK
JournalMethods Mol Biol
PubMed ID9664375
Targeting of small unilamellar liposomes to the galactose receptor in vivo.
AuthorsGregoriadis G, Senior J
JournalBiochem Soc Trans
PubMed ID6327432
Coupling of monoclonal antibodies with fluorophores.
AuthorsHaugland RP
JournalMethods Mol Biol
PubMed ID7550684
The use of phycobiliproteins as fluorescent labels in immunoassay.
AuthorsKronick MN
JournalJ Immunol Methods
PubMed ID3528294
Cross-linking techniques.
AuthorsBäumert HG, Fasold H
JournalMethods Enzymol
PubMed ID2546017
Preparation of fluorescent, cross-linking, and biotinylated calmodulin derivatives and their use in studies of calmodulin-activated phosphodiesterase and protein phosphatase.
AuthorsKincaid RL, Billingsley ML, Vaughan M
JournalMethods Enzymol
PubMed ID2842624
Protein labeling with fluorescent probes.
AuthorsHolmes KL, Lantz LM
JournalMethods Cell Biol
PubMed ID11060842
Antibody-coated liposomes as a particulate solid phase for immunoassays. Measurement of urinary 'micro-albumin'.
AuthorsFrost SJ, Firth GB, Chakraborty J
JournalJ Immunol Methods
PubMed ID2254667
A novel use of liposomes as a solid phase material achieving separation in immunoassays is described. Antibody-coated liposomes were prepared and used as a particulate solid phase in a radioimmunoassay procedure for urinary albumin. The assay was compared to a liquid phase albumin radioimmunoassay. The potential benefits of liposomes over ... More
Preparation of reproducible alkaline phosphatase-antibody conjugates for enzyme immunoassay using a heterobifunctional linking agent.
AuthorsJeanson A, Cloes JM, Bouchet M, Rentier B
JournalAnal Biochem
PubMed ID2847583
Conjugates of alkaline phosphatase (AP) and mouse monoclonal immunoglobulins G (IgG) were prepared by means of the heterobifunctional linker, N-succinimidyl 3-(2-pyridyldithio)-propionate. The efficiency of such conjugates can be improved by optimizing the degree of substitution of IgG and AP. We have determined conditions yielding better performing conjugates than those synthesized ... More
In migrating fibroblasts, recycling receptors are concentrated in narrow tubules in the pericentriolar area, and then routed to the plasma membrane of the leading lamella.
AuthorsHopkins CR, Gibson A, Shipman M, Strickland DK, Trowbridge IS
JournalJ Cell Biol
PubMed ID7515888
By following the intracellular processing of recycling transferrin receptors and the selective sorting of a-2 macroglobulin in chick embryo fibroblasts, we have shown that the concentration of 60 nm diam tubules which surrounds the centrioles represents a distal compartment on the recycling pathway. In migrating cells transferrin receptor tracers can ... More
Direct determination of molecular ratios of peptides coupled via N-succinimidyl 3-(2-pyridyldithio)propionate to carrier proteins using high performance liquid chromatography.
AuthorsBläsi U, Linke RP, Lubitz W
JournalJ Immunol Methods
PubMed ID3351310
A simple, rapid and reproducible method is presented for direct determination of the substitution ratio of a carrier protein with a synthetic nonradioactively labeled peptide. The peptide was covalently linked by a thiol group of a cysteine residue to the immunogenic carrier protein using the heterobifunctional reagent N-succinimidyl 3-(2-pyridyldithio)propionate. The ... More
Determination of lysozyme in serum, urine, cerebrospinal fluid and feces by enzyme immunoassay.
AuthorsBrouwer J, van Leeuwen-Herberts T, Otting-van de Ruit M
JournalClin Chim Acta
PubMed ID6383662
Conjugates of human lysozyme and horseradish peroxidase (HRP) were prepared by means of the heterobifunctional reagent N-succinimidyl 3-(2-pyridyldithio) propionate. A conjugate containing 2 mol HRP/mol lysozyme was isolated by gel filtration and used as a labeled antigen in competitive enzyme immunoassays, in which anti-lysozyme rabbit IgG had been bound to ... More
Endocytosis of liposomes bound to cell surface proteins measured by flow cytofluorometry.
AuthorsTruneh A, Mishal Z, Barbet J, Machy P, Leserman LD
JournalBiochem J
PubMed ID6615464
A new technique for the quantification of cellular receptor-mediated endocytosis has been developed based on the analysis by flow cytometry of ligand-bearing liposomes containing the fluorochrome carboxyfluorescein. Carboxyfluorescein encapsulated at high concentrations in protein A-bearing liposomes is self-quenched. Binding and internalization of such liposomes by cells via antibodies directed towards ... More
Enzyme-linked fluorescence immunoassays using beta-galactosidase and antibodies covalently bound to polystyrene plates.
AuthorsNeurath AR, Strick N
JournalJ Virol Methods
PubMed ID6795220
A solid-phase enzyme-linked immunoassay using a fluorogenic substrate (4-methylumbelliferyl-beta-D-galactopyranoside) was developed. Antibodies were covalently linked to glutaraldehyde-activated 96-well aminopolystyrene plates. Antigens from test samples were adsorbed to the solid phase and detected using antibodies conjugated with E. coli beta-galactosidase. Glutaraldehyde, N-succinimidyl-3-(2-pyridyldithio)-propionate or N-succinimidyl-6-(4-azido-2-nitrophenylamino)-hexanoate were used as linkers between antibodies and ... More
A simple spectrophotometric determination of solid supported amino groups.
AuthorsNgo TT
JournalJ Biochem Biophys Methods
PubMed ID3734326
A simple spectrophotometric method for the quantitative determination of solid phase supported amino groups is described. The method involves reacting the solid support with an excess of activated acylating agent, N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP) and an efficient acylation catalyst, 4-dimethylaminopyridine, and after thoroughly removing the unreacted SPDP, the solid support is reacted ... More
Immunoassay techniques with fluorescent phycobiliprotein conjugates.
AuthorsKronick MN, Grossman PD
JournalClin Chem
PubMed ID6883673
We describe immunoassay techniques that take advantage of the novel properties of phycobiliprotein fluorescent dyes. These dyes, which can be isolated from a wide variety of algae, exhibit extremely high absorptivities, high quantum efficiencies, and excitation and emission bands across the visible spectrum. These stable, hydrophilic proteins can easily be ... More
Novel protein transfection of primary rat cortical neurons using an antibody that penetrates living cells.
AuthorsWeisbart RH, Baldwin R, Huh B, Zack DJ, Nishimura R
JournalJ Immunol
PubMed ID10820286
An Ab-based system to deliver functional proteins into neurons was developed using the murine mAb, mAb 3E10. This was achieved by covalently conjugating catalase to the Ab so that the conjugate retained high activity for the degradation of hydrogen peroxide. Three-dimensional fluorescence microscopy was used to demonstrate penetration of the ... More
Preparation and characterization of xanthine oxidase-antibody and -hapten conjugates for use in sensitive chemiluminescent immunoassays.
AuthorsFert V, Baret A
JournalJ Immunol Methods
PubMed ID2391429
Methods have been developed to label haptens or antibodies with xanthine oxidase for use in chemiluminescent enzyme immunoassays. We have optimised coupling reactions involving the use of heterobifunctional cross-linkers, the introduction of sulfhydryl groups and the utilization of accessible cysteine residues on the native enzyme. The versatility of xanthine oxidase ... More
Fluorescent tandem phycobiliprotein conjugates. Emission wavelength shifting by energy transfer.
AuthorsGlazer AN, Stryer L
JournalBiophys J
PubMed ID6414547
A fluorescent tandem phycobiliprotein conjugate with a large Stokes shift was prepared by the covalent attachment of phycoerythrin to allophycocyanin. The efficiency of energy transfer from phycoerythrin to allophycocyanin in this disulfide-linked conjugate was 90%. A distinctive feature of this phycocyanin conjugate is the wide separation between the intense absorption ... More
Vitamin B12 mediated oral delivery systems for granulocyte-colony stimulating factor and erythropoietin.
AuthorsRussell-Jones GJ, Westwood SW, Habberfield AD
JournalBioconjug Chem
PubMed ID7578366
As a prelude to the development of orally active erythropoietin (EPO) and granulocyte-colony stimulating factor (G-CSF), conjugates have been formed between these molecules and vitamin B12. During the formation of these conjugates intramolecular cross-linking of the proteins was avoided by the use of hydrazidyl derivatives of vitamin B12. A potentially ... More
Liposome immune lysis assay (LILA): a simple method to measure anti-protein antibody using protein antigen-bearing liposomes.
AuthorsIshimori Y, Yasuda T, Tsumita T, Notsuki M, Koyama M, Tadakuma T
JournalJ Immunol Methods
PubMed ID6549183
A new simple immunoassay technique using immune lysis of liposomes was developed to measure antibody against protein antigens. Multilamellar liposomes were composed of dipalmitoylphosphatidylcholine, cholesterol and phosphatidylethanolamine substituted with the hetero-bifunctional cross-linking reagent N-hydroxysuccinimidyl 3-(2-pyridyldithio)propionate (SPDP). The protein antigen (human IgG) was coupled to these liposomes after treatment with SPDP ... More
Detection of low-density cell-surface molecules using biotinylated fluorescent microspheres.
AuthorsWojchowski DM, Sytkowski AJ
JournalBiochim Biophys Acta
PubMed ID3964705
Biotinylated fluorescent microspheres have been developed as a reagent for studying antigens and receptors expressed at the cell surface. Labeling of antigen or receptor was accomplished by crosslinking biotinylated microspheres through streptavidin to corresponding biotinylated antibodies or ligands. Detection of labeled cells by flow microfluorimetry provided an extremely sensitive means ... More
Near-infrared fluorescence lifetime assay for serum glucose based on allophycocyanin-labeled concanavalin A.
AuthorsMcCartney LJ, Pickup JC, Rolinski OJ, Birch DJ
JournalAnal Biochem
PubMed ID11355853
We describe an assay scheme for glucose based on fluorescence resonance energy transfer (FRET) between concanavalin A (con A), labeled with the near-infrared fluorescent protein allophycocyanin (APC) as donor, and dextran labeled with malachite green (MG) as acceptor. Glucose competitively displaces dextran-MG and leads to reduction in FRET, assessed by ... More
Rolling properties of rGPIbalpha-conjugated phospholipid vesicles with different membrane flexibilities on vWf surface under flow conditions.
AuthorsTakeoka S, Teramura Y, Okamura Y, Tsuchida E, Handa M, Ikeda Y
JournalBiochem Biophys Res Commun
PubMed ID12176049
The recombinant fragment of the platelet membrane glycoprotein, rGPIbalpha, was conjugated to phospholipid vesicles with the average diameter of ca. 1 microm using N-succinimidyl 3-(2-pyridyldithio) propionate (SPDP). We used five kinds of rGPIbalpha-vesicles with different fluorescent anisotropies of 1,6-diphenyl-1,3,5-hexatriene (DPH) to study the rolling properties of the vesicles on the ... More
Liposome immunoassay by long-lived fluorescence detection.
AuthorsOkabayashi Y, Ikeuchi I
JournalAnalyst
PubMed ID9764512
Immunoassay by fluorescence energy transfer from a europium chelate in liposome to allophycocyanin (APC) was demonstrated. Streptavidin or antibody to biotin was bonded to the liposome containing the europium chelate of 2-naphthoyltrifluoroacetone in the bilayer. When the biotin bonded to APC (APC-BT) was added to the prepared liposomes and the ... More
Synthesis and applications of a new poly(ethylene glycol) derivative for the crosslinking of amines with thiols.
AuthorsHaselgrübler T, Amerstorfer A, Schindler H, Gruber HJ
JournalBioconjug Chem
PubMed ID7632794
A heterobifunctional crosslinker was synthesized from a diamine derivative of poly(ethylene glycol) (PEG, average molecular weight 800 Da), with the functional groups 2-(pyridyldithio)propionyl (PDP) and N-hydroxysuccinimide ester (NHS). The crosslinker can be used for linkage of two different proteins for which a suitable protocol is presented, explified by crosslinking of ... More
Development of sensitive immunoassays for detection of antibodies against hepatitis B surface antigen.
AuthorsIonescu-Matiu I, Sanchez Y, Fields HA, Dreesman GR
JournalJ Virol Methods
PubMed ID6403563
Three micro solid phase immunoassays (a micro-SPRIA and two ELISA techniques) were developed and tested for the detection of anti-HBs antibodies. Two different crosslinkers (glutaraldehyde and N-succinimidyl 3-(2-pyridyldithio) propionate) were used to couple a goat anti-mouse IgG reagent to alkaline phosphatase for use as enzyme-labeled probes in the two ELISA ... More
Conjugation of plasminogen activators and fibrin-specific antibodies to improve thrombolytic therapeutic agents.
AuthorsHayzer DJ, Lubin IM, Runge MS
JournalBioconjug Chem
PubMed ID1790168
Here we have reviewed chemical and recombinant approaches to the construction of hybrid molecules that combine a "targeting" antibody and an "effector" enzyme activity. There are advantages and disadvantages to both chemical and recombinant methods, and one goal of this review has been to elucidate these so that the appropriate ... More
Preparation of protein A-peroxidase monoconjugate using a heterobifunctional reagent, and its use in enzyme immunoassays.
AuthorsPain D, Surolia A
JournalJ Immunol Methods
PubMed ID6265559
Protein A-peroxidase monoconjugate was prepared in solution using a heterobifunctional reagent, N-succinimidyl-3-(2-pyridyldithio)-propionate. The yield of the monoconjugate was much higher than that obtained with current methods. An immunoassay method was developed in which protein A-peroxidase monoconjugate served as a universal tool. Protein A-peroxidase monoconjugate was taken up by IgG molecules ... More
Selective killing of target cells by antibody-ricin A chain or antibody-gelonin hybrid molecules: comparison of cytotoxic potency and use in immunoselection procedures.
AuthorsColombatti M, Nabholz M, Gros O, Bron C
JournalJ Immunol
PubMed ID6139402
Ricin A chain and gelonin, two plant proteins that can powerfully reduce the protein synthetic capacity of ribosome preparations, were covalently coupled to a monoclonal anti-Thy-1.2 antibody. Two conjugates were prepared by using N-succinimidyl-3-2(-pyridyldithio)propionate, which generates a disulfide linkage between the component molecules. Both conjugates specifically killed Thy-1.2 antigen-bearing EL-4 ... More
Preparation of enzyme-conjugated DNA probe and application to the universal probe system.
AuthorsNakagami S, Matsunaga H, Oka N, Yamane A
JournalAnal Biochem
PubMed ID1789434
A general procedure for the cross-linking of enzyme to DNA has been developed for use as a nonradioactive probe. In this method, DNA is transaminated with diaminopropane to introduce primary amino groups into the cytosine residues. Then the amino groups are converted to thiol groups using a heterobifunctional cross-linker. The ... More
Chemically optimized antimyosin Fab conjugates with chelating polymers: importance of the nature of the protein-polymer single site covalent bond for biodistribution and infarction localization.
Murine antimyosin Fab fragment was conjugated with 111In-labeled N-terminal-modified DTPA-polylysine using three bifunctional reagents: N-hydroxysuccinimide esters of 3-(2-pyridyldithio)propionic acid (SPDP conjugate), 4-(maleimidomethyl)cyclohexanecarboxylic acid (SMCC conjugate) and bromoacetic acid (BrAc conjugate) for potential localization of experimental myocardial infarction. Using various antibody preparations and a rabbit acute myocardial infarction model the following ... More
The use of maleimidocaproyloxysuccinimide to prepare malarial peptide carrier immunogens. Immunogenicity of the linking region.
AuthorsJones GL, Edmundson HM, Spencer L, Gale J, Saul A
JournalJ Immunol Methods
PubMed ID2478635
Malarial peptides synthesized with an added N terminal cysteine were conjugated to purified diphtheria toxoid (DT) protein using the bifunctional reagent maleimidocaproyloxysuccinimide (MCS). The molar ratio of peptide to carrier was determined by subtractive sulphydryl titration and confirmed by sodium dodecyl sulphate (SDS) electrophoresis. For enzyme-linked immunoabsorbent (ELISA) analysis of ... More