ELISA: Protocols

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Overview

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.

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

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

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

1. Abatacept
2. Adalimumab
3. Atezolizumab
4. Avelumab
5. Avelumab  (TZA041, TZA040P)
6. Belatacept
7. Bevacizumab
8. Brentuximab vedotin
9. Cemiplimab
10. Cetuximab
11. Daratumumab
12. Denosumab
13. Dupilumab
14. Durvalumab
15. Eculizumab
16. Etanercept
17. Evolocumab
18. Golimumab
19. Infliximab
20. Ipilimumab
21. Natalizumab
22. Nivolumab
23. Obinutuzumab
24. Ocrelizumab
25. Ocrelizumab  (TZA044, TZA045)
26. Omalizumab
27. Palivizumab
28. Panitumumab
29. Pembrolizumab
30. Pertuzumab
31. Rituximab
32. Secukinumab
33. Tocilizumab
34. Trastuzumab (HCA166, HCA168)
35. Trastuzumab  (HCA169, HCA176P)
36. Trastuzumab (HCA169, HCA177P)
37. Trastuzumab (HCA169, HCA270 IgG4)
38. Ustekinumab  (HCA208, HCA210)
39. Ustekinumab  (HCA209, HCA210)
40. Vedolizumab 

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

ADA Bridging ELISA

Type 1 anti-idiotypic antibodies are neutralizing and fully human; in immunoglobulin format they can be used as a control or calibrator as a surrogate for human ADAs

1. Abatacept
2. Adalimumab (IgG1)
3. Adalimumab (IgE)
4. Atezolizumab
5. Avelumab
6. Belatacept
7. Bevacizumab
8. Brentuximab vedotin
9. Cemiplimab
10. Certolizumab pegol
11. Cetuximab
12. Daratumumab
13. Denosumab
14. Dupilumab
15. Durvalumab
16. Eculizumab
17. Etanercept
18. Evolocumab
19. Infliximab
20. Ipilimumab
21. Natalizumab
22. Nivolumab
23. Obinutuzumab
24. Ocrelizumab
25. Omalizumab
26. Palivizumab
27. Panitumumab
28. Pembrolizumab
29. Pertuzumab
30. Ranibizumab
31. Rituximab
32. Secukinumab
33. Tocilizumab
34. Trastuzumab
35. Ustekinumab
36. Vedolizumab

 

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.

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. Atezolizumab  (TZA013)
3. Atezolizumab  (TZA013P)
4. Avelumab
5. Avelumab  (TZA042P)
6. Golimumab
7. Durvalumab
8. Ipilimumab  (HCA331)
9. Ipilimumab (TZA001P)
10. Omalizumab
11. Ranibizumab (HCA304, HCA306, HCA307)
12. Ranibizumab (HCA316P)
13. Secukinumab
14. Trastuzumab