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. 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

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. Belatacept
6. Bevacizumab
7. Brentuximab vedotin
8. Cemiplimab
9. Certolizumab pegol
10. Cetuximab
11. Daratumumab
12. Denosumab
13. Dupilumab
14. Durvalumab
15. Eculizumab
16. Etanercept
17. Evolocumab
18. Infliximab
19. Ipilimumab
20. Natalizumab
21. Nivolumab
22. Omalizumab
23. Palivizumab
24. Panitumumab
25. Pembrolizumab
26. Pertuzumab
27. Ranibizumab
28. Rituximab
29. Secukinumab
30. Tocilizumab
31. Trastuzumab
32. Ustekinumab
33. 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. 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