Hello, does anyone working with antibody fragments (Fab and Fc) conjugation or knows the chemical structure/functional groups related to conjugation? If yes,kindly let me know, I have a query. Thank you.
You can use biotinylation reagent ( for example, https://www.thermofisher.com/order/catalog/product/21343 )
Article Characterization for Binding Complex Formation with Site-Dir...
The options for conjugation will depend to some extent on how the fragments were generated. For example, if Fab has been produced from IgG enzymatically (via pepsin and Fab'2 reduction) you will have thiol groups which are highly reactive (and located away from the antigen binding areas) and these can be conjugated to maleimide or iodoacetyl derivatives. If Fab has been generated using papain then the thiols will be absent. If small fragments are recombinantly expressed it is common to add a terminal cysteine for the purpose of conjugation. However, regrdless of how the fragments have been generated you will always have plenty of lysine residues and there are numerous methods for conjugating labels to those.
Thank you Mr. Nick Gee . I will do Fab'2 reduction by pepsin. Does Fab'2 contain thiol group or S-S bond at the end of two Fab joint? As I see, it contains S-S bond but I want to keep both Fab fragments together and conjugate Fc with Fab'2. Without separating two Fab from Fab'2 ,can I make any free thiol group to conjugate with another thiol group in Fc? In this case, Fc might not have free SH. How I can proceed please guide me, Sir. This is new field for me but I must do it.
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