Is it necessary to have pilot study for antibody fragmentation?
Because individual antibodies can have different susceptibilities to the enzyme, sometimes, if not carefully done, enzyme can cause complete degradation fot he antibody to peptides. Cysteine is recommended for reducing IgM to IgMs because stronger reducing agents can cleave the disulfide bonds between the heavy chains within the subunit. Because alkylation is necessary to prevent spontaneous reassociation of the subunits, the single-step reduction and alkylation offered by cysteine is beneficial. Individual antibodies can have different susceptibilities to cysteine; thus, a pilot fragmentation experiment is recommended for optimizing conditions.
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07/02/2012