Analysis Antibody Binding Affinity and Specificity of Biopharmaceutical Compounds by Enzyme-linked Immunosorbent Assay (ELISA) (CAT#: STEM-B-0387-CJ)

Introduction

Biological activity is, besides the safety of the therapy, the most important aspect of a drug product or therapy. Thus, a major goal of formulation development is to maintain the functionality of the therapeutic compound inside its formulation throughout its shelf-life.

The biologic functions are isotype- and subclass-dependent. In comparison with specificity, affinity of an antibody is a measure of the strength of the binding between antibody and antigen, such that a low-affinity antibody binds weakly and high-affinity antibody binds firmly.

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Principle Applications Procedure Materials Notes Related Products

Principle

ELISA works on the principle that specific antibodies bind the target antigen and detect the presence and quantity of antigens binding. In order to increase the sensitivity and precision of the assay, the plate must be coated with antibodies with high affinity.

Applications

Biopharmaceutica

Procedure

1. Attachment of capture antibody specific to target protein to a microplate.
2. Addition of standards and samples containing unknown amount of the target protein which binds to the capture antibody.
3. Washing to remove unbound substances.
4. Addition of a detection antibody that binds to the immobilized target protein
5. Washing away excess detection antibody and addition of HRP conjugate.
6. Addition of HRP substrate for indirect detection of bound protein.

Materials

• Sample: Peptides, Proteins, Vaccines, Virus-like particles
• Equipment: ELISA kits

Notes

The ELISA plates have the reaction products immunosorbed on the solid phase, which is part of the plate, and so are not easily reusable.

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