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[rick112]

hi

what is the method generally used for F(ab’)2 Preparation ?? (i have come across protocols for both pepsin and ficin )

also how do you purify F(ab’)2 Preparation ?

{i am intertesed in obtaining only F(ab’)2 segment of IgG1 antibody }

thanks

[BioMiha]

In theory you can generate the F(ab')2 fragment with pepsin, although I don't have direct experience with that. In our hands ficin did not work very well.
I have done many Fab preparations, digesting the IgG1 with papain. For purification the best way is to remove the undigested Abs with protein A or G and then maybe do another purification step - I use an affinity column that I prepared myself with anti-Fab antibodies.