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.

The Thermo Scientific Pierce LAL Chromogenic Endotoxin Quantitation Kit measures the amount of endotoxin in a protein, peptide or antibody sample using the LAL assay. Designed for up to 50 samples, this quantitation kit is 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 0.1 EU/mL. Protein and antibody samples can be assayed in as few as 30 minutes.

  • Sensitive—detect as little as 0.1 EU/mL
  • Fast—perform assay in 30 minutes
  • Minimal sample—assay requires only 10 μL of a protein sample
  • Accurate—E. coli O111:B4 standard in each kit enables accurate endotoxin quantitation
  • Versatile—405 nm absorbance reading is compatible with common ELISA plate readers

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. Clean-up proteins of various molecular weights and isoelectric points with minimal sample. 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%


Learning resources

Application Notes