How can I conjugate DNP to an IgG antibody in a 1:5 molar ratio?

07 January 2025 0 6K Report

I want to conjugate DNP (2,4-dinitrophenyl, molecular weight: 394 Da) to an IgG antibody (molecular weight: 150 kDa) in a 1:5 molar ratio (1 antibody to 5 DNP molecules).

I would like to know:

How to calculate the amount of DNP required for the conjugation based on the desired molar ratio?

What are the best chemical methods or protocols for conjugating DNP to IgG while preserving the antibody's structure and functionality?

Are there any recommended purification techniques to remove unbound DNP after conjugation?

How can I confirm the success of the conjugation (e.g., spectrophotometry, mass spectrometry, or other methods)?

I would appreciate any suggestions, protocols, or tips to optimize this process. Thank you!

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