Endotoxin Quantitation and Removal

Endotoxins are heat-stable lipopolysacchride (LPS) molecules associated with the outer membranes of certain gram negative bacteria. When these bacterial cells die, their cell membranes rupture and endotoxins are released into the surrounding environment. Endotoxins are frequent contaminants of protein solutions derived from bioproduction and are toxic to cells grown in tissue culture.

Choose the right endotoxin quantitation or removal product

We offer two kits that accurately detect the amount of endotoxin in a protein, peptide or antibody sample using an endpoint amebocyte lysate assay. These quantitation kits are ideal for processing a few samples at a time compared to other high-throughput quantitation kits. The endotoxin concentration in a sample is measured via a chromogenic signal generated in the presence of endotoxins. Samples can be measured on a microplate absorbance reader at 405 nm. A standard curve is created using the E. coli endotoxin standard included with each kit to calculate endotoxin levels as low as 0.01 EU/mL in 16-36 minutes, depending on the sensitivity needed.

  • Sensitive—detect as little as 0.01 EU/mL
  • Fast—perform assays in 16-36 minutes
  • Minimal sample—assay requires only 10 μL of a protein sample
  • AccurateE. coli O111:B4 standard in each kit enables accurate endotoxin quantitation
  • Versatile—405 nm absorbance reading is compatible with common ELISA plate readers

 

Choose the right endotoxin quantitation kit

  Pierce LAL Chromogenic Endotoxin Quantitation Kit Pierce Chromogenic Endotoxin Quant Kit
Sensitivity range 0.1 - 1.0 0.01 - 0.1  (low)
 0.1 - 1.0
Assay time - high range (min) 16 20
Assay time - low range (min) NA 36
β-glucan (10 ng/mL) NA Compatible
Pack sizes 50-reaction kit 30-, 60- and 240-reaction kits
  Order now Order now

 

Standard Curves

Standard curves for Pierce endotoxin quantitation kits

Figure 1. Standard curves for Pierce endotoxin quantitation kits. The constructed endotoxin standard curves shows exceptional linearity with r2 =0.99. Pierce Chromogenic Endotoxin Quant Kit has a lower range standard curve between 0.01–0.1 EU/mL. The standard curve, 0.1–1.0 EU/mL represents 14-minute incubation with endotoxin standards and amebocyte lysate followed by 6-minute incubation with the chromogenic substrate. For the lower range, 30-minute incubation with endotoxin standards and amebocyte lysate was followed by 6-minute incubation with the chromogenic substrate. Consistency and reproducibility (n=17; CV=3%) is shown for the low range standards (0.01–0.1 EU/mL).

Thermo Scientific Pierce High Capacity Endotoxin Removal Resin selectively binds and removes endotoxins from protein, peptide and antibody samples using a modified ε-poly-L-lysine [poly(ε-lysine)] affinity ligand. Endotoxin levels in biological samples are reduced by up to 99% in as fast as 1 hour using our spin column format, and protein recovery is ≥ 85%. Pierce High Capacity Endotoxin Removal Resin is available as a slurry to pack a custom column or in convenient pre-packed, single-use spin columns optimized for different sample volumes.

  • High capacity—bind up to 2,000,000 EU/mL to eliminate >99% of endotoxins
  • Durable—reuse resin up to 10 times
  • Selective—recover ≥85% of your protein sample
  • High performance—complies with FDA guidelines by reducing final EU concentration to <5 EU/mL
  • Fast—our spin column format enables endotoxin depletion typically within 1 hour
  • Clean—single-use spin columns avoid cross contamination of samples
  • Optimized—spin columns are optimized for different sample volumes
  • Economical—large-volume discounts available

 

Choose the right endotoxin removal product

 
Format Spin column Spin column Spin column Loose resin
Column size (mL) 0.25 0.5 1 Varies
Sample volume (mL) 0.5 - 1 1.0 - 4.0 2.0 - 10 Varies
Order 5 columns 5 columns 5 columns 10 mL
25 columns 25 columns 100 mL
250 mL

Binding capacity

Table 1. Efficiently remove endotoxins from heavily contaminated samples. Thermo Scientific Pierce High Capacity Endotoxin Removal Spin Columns, 0.5 mL, were challenged for 1 hour with protein (2 mL of 1 mg/mL BSA) containing 5,000 to 500,000 EU/mL from E. coli strain 055:B5. Even at the highest endotoxin levels clean-up was >98%.

Initial endotoxin concentration (EU/mL)* Final endotoxin concentration (EU/mL) Endotoxin removal effi ciency Protein recovery
5,000 <1 99.98% >90%
12,500 <1 99.99% >90%
25,000 1.26 99.99% >90%
50,000 7.1 99.99% >90%
250,000 32 99.98% >90%
500,000 9,600 99.08% >90%

Protein compatibility

Table 2. Endotoxin removal and protein recovery of various proteins at different molecular weights and isoelectric points. Pierce High Capacity Endotoxin Removal Spin Columns, 0.5 mL, were challenged for 1 hour with different proteins of various molecular weight and charge. In all samples endotoxin removal was >99% and protein recovery was >85%.

Protein (1 mg/mL) Molecular weight (Da) Isoelectric point (pI) Initial endotoxin concentration (EU/mL) Final endotoxin concentration (EU/mL) Endotoxin removal efficiency Protein recovery
Cytochrome C 12,000 10.6 10,000 1.35 >99.9% 86%
Myoglobin 17,000 6.8 10,000 3.68 >99.9% 87%
Bovine serum albumin (BSA) 66,000 4.9 10,000 0.80 >99.9% 85%
Bovine gamma globulin (BGG) 150,000 7.4 10,000 4.60 >99.9% 92%
>99.9%
>99.9%

We offer two kits that accurately detect the amount of endotoxin in a protein, peptide or antibody sample using an endpoint amebocyte lysate assay. These quantitation kits are ideal for processing a few samples at a time compared to other high-throughput quantitation kits. The endotoxin concentration in a sample is measured via a chromogenic signal generated in the presence of endotoxins. Samples can be measured on a microplate absorbance reader at 405 nm. A standard curve is created using the E. coli endotoxin standard included with each kit to calculate endotoxin levels as low as 0.01 EU/mL in 16-36 minutes, depending on the sensitivity needed.

  • Sensitive—detect as little as 0.01 EU/mL
  • Fast—perform assays in 16-36 minutes
  • Minimal sample—assay requires only 10 μL of a protein sample
  • AccurateE. coli O111:B4 standard in each kit enables accurate endotoxin quantitation
  • Versatile—405 nm absorbance reading is compatible with common ELISA plate readers

 

Choose the right endotoxin quantitation kit

  Pierce LAL Chromogenic Endotoxin Quantitation Kit Pierce Chromogenic Endotoxin Quant Kit
Sensitivity range 0.1 - 1.0 0.01 - 0.1  (low)
 0.1 - 1.0
Assay time - high range (min) 16 20
Assay time - low range (min) NA 36
β-glucan (10 ng/mL) NA Compatible
Pack sizes 50-reaction kit 30-, 60- and 240-reaction kits
  Order now Order now

 

Standard Curves

Standard curves for Pierce endotoxin quantitation kits

Figure 1. Standard curves for Pierce endotoxin quantitation kits. The constructed endotoxin standard curves shows exceptional linearity with r2 =0.99. Pierce Chromogenic Endotoxin Quant Kit has a lower range standard curve between 0.01–0.1 EU/mL. The standard curve, 0.1–1.0 EU/mL represents 14-minute incubation with endotoxin standards and amebocyte lysate followed by 6-minute incubation with the chromogenic substrate. For the lower range, 30-minute incubation with endotoxin standards and amebocyte lysate was followed by 6-minute incubation with the chromogenic substrate. Consistency and reproducibility (n=17; CV=3%) is shown for the low range standards (0.01–0.1 EU/mL).

Thermo Scientific Pierce High Capacity Endotoxin Removal Resin selectively binds and removes endotoxins from protein, peptide and antibody samples using a modified ε-poly-L-lysine [poly(ε-lysine)] affinity ligand. Endotoxin levels in biological samples are reduced by up to 99% in as fast as 1 hour using our spin column format, and protein recovery is ≥ 85%. Pierce High Capacity Endotoxin Removal Resin is available as a slurry to pack a custom column or in convenient pre-packed, single-use spin columns optimized for different sample volumes.

  • High capacity—bind up to 2,000,000 EU/mL to eliminate >99% of endotoxins
  • Durable—reuse resin up to 10 times
  • Selective—recover ≥85% of your protein sample
  • High performance—complies with FDA guidelines by reducing final EU concentration to <5 EU/mL
  • Fast—our spin column format enables endotoxin depletion typically within 1 hour
  • Clean—single-use spin columns avoid cross contamination of samples
  • Optimized—spin columns are optimized for different sample volumes
  • Economical—large-volume discounts available

 

Choose the right endotoxin removal product

 
Format Spin column Spin column Spin column Loose resin
Column size (mL) 0.25 0.5 1 Varies
Sample volume (mL) 0.5 - 1 1.0 - 4.0 2.0 - 10 Varies
Order 5 columns 5 columns 5 columns 10 mL
25 columns 25 columns 100 mL
250 mL

Binding capacity

Table 1. Efficiently remove endotoxins from heavily contaminated samples. Thermo Scientific Pierce High Capacity Endotoxin Removal Spin Columns, 0.5 mL, were challenged for 1 hour with protein (2 mL of 1 mg/mL BSA) containing 5,000 to 500,000 EU/mL from E. coli strain 055:B5. Even at the highest endotoxin levels clean-up was >98%.

Initial endotoxin concentration (EU/mL)* Final endotoxin concentration (EU/mL) Endotoxin removal effi ciency Protein recovery
5,000 <1 99.98% >90%
12,500 <1 99.99% >90%
25,000 1.26 99.99% >90%
50,000 7.1 99.99% >90%
250,000 32 99.98% >90%
500,000 9,600 99.08% >90%

Protein compatibility

Table 2. Endotoxin removal and protein recovery of various proteins at different molecular weights and isoelectric points. Pierce High Capacity Endotoxin Removal Spin Columns, 0.5 mL, were challenged for 1 hour with different proteins of various molecular weight and charge. In all samples endotoxin removal was >99% and protein recovery was >85%.

Protein (1 mg/mL) Molecular weight (Da) Isoelectric point (pI) Initial endotoxin concentration (EU/mL) Final endotoxin concentration (EU/mL) Endotoxin removal efficiency Protein recovery
Cytochrome C 12,000 10.6 10,000 1.35 >99.9% 86%
Myoglobin 17,000 6.8 10,000 3.68 >99.9% 87%
Bovine serum albumin (BSA) 66,000 4.9 10,000 0.80 >99.9% 85%
Bovine gamma globulin (BGG) 150,000 7.4 10,000 4.60 >99.9% 92%
>99.9%
>99.9%

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