The enzyme-linked immunosorbent assay (ELISA) is a frequently used application which determines the concentration of an analyte in solution. The ELISA method is straightforward and rapid. With automated equipment the assay is easily scalable for set up in high-throughput and can handle a large number of samples in parallel, making it a popular choice for the evaluation of research and diagnostic targets.

ELISA Protocols

General ELISA Protocols

Standard methods for different types of ELISA in 96-well microtiter plate format, including recommended buffers.

  1. Indirect
  2. Competition (Inhibition)
  3. Direct ELISA with streptavidin-biotin detection
  4. Sandwich ELISA with streptavidin-biotin detection
  5. Sandwich ELISA with direct detection

Protocols for use with Anti-Biotherapeutic Antibodies

Bio-Rad’s anti-biotherapeutic antibodies are ideal for the measurement of monoclonal antibody drugs and biosimilar products in PK and ADA assays.

We have developed a range of highly specific, recombinant anti-idiotypic antibodies with different binding modes and properties, enabling the development of bridging ELISA and antigen capture ELISA. Protocols are available for each type of PK assay and ADA assay for many of the anti-biotherapeutic products in our catalog.

PK and ADA Bridging ELISA Protocols

Bridging ELISA is a special case of a sandwich ELISA in which a dimeric or oligomeric antigen (most often an antibody in a sample) is detected by a capture and detection antibody. The antigen bridges the two specific antibodies. Bridging ELISAs are most frequently used for the detection of IgG, for example in pharmacokinetic (PK) or anti-drug antibody (ADA) assays.

PK Bridging ELISA measuring free drug protocols

PK Bridging ELISA measuring free drug

Type 1 anti-idiotypic antibodies are inhibitory and are used to detect free drug

  1. Abatacept
  2. Adalimumab
  3. Atezolizumab
  4. Belatacept
  5. Bevacizumab
  6. Brentuximab vedotin
  7. Cemiplimab
  8. Cetuximab
  9. Daratumumab
  10. Denosumab
  11. Dupilumab
  12. Durvalumab
  13. Eculizumab
  14. Etanercept
  15. Evolocumab
  16. Golimumab
  17. Infliximab
  18. Ipilimumab
  19. Natalizumab
  20. Nivolumab
  21. Omalizumab
  22. Palivizumab
  23. Panitumumab
  24. Pembrolizumab
  25. Pertuzumab
  26. Rituximab
  27. Secukinumab
  28. Tocilizumab
  29. Trastuzumab (HCA166, HCA168)
  30. Trastuzumab  (HCA169, HCA176P)
  31. Trastuzumab (HCA169, HCA177P)
  32. Trastuzumab (HCA169, HCA270 IgG4)
  33. Ustekinumab  (HCA208, HCA210)
  34. Ustekinumab  (HCA209, HCA210)
  35. Vedolizumab 
PK Bridging ELISA measuring total drug protocols

PK Bridging ELISA measuring total drug

Type 2 anti-idiotypic antibodies are non-inhibitory and enable the measurement of total drug – free, partially and fully bound

  1. Cetuximab
  2. Dupilumab
  3. Etanercept
  4. Golimumab
  5. Infliximab
  6. Tocilizumab

Antigen Capture ELISA Protocols

A third type of antibody that only detects drug bound to its target antigen is available for some of our specificities.

Protocols: PK Antigen Capture ELISA measuring bound drug exclusively

PK Antigen Capture ELISA measuring bound drug exclusively

A Type 3 antibody is used in an antigen capture ELISA to detect bound drug exclusively and avoids the bridging ELISA format

  1. Adalimumab
  2. Golimumab
  3. Durvalumab
  4. Ipilimumab (HCA331)
  5. Ipilimumab (TZA001P
  6. Omalizumab
  7. Ranibizumab (HCA304, HCA306, HCA307
  8. Ranibizumab (HCA316P)
  9. Secukinumab
  10. Secukinumab (HCA374)
  11. Trastuzumab

Technical Support

Technical Support