Any techniques to assess if the antibody maintain the original conformation after break one interchain disulfide bond? Thank you.
Depends what you are trying to achieve. If the concern is over loss of binding function, then a simple Biacore experiment can make sure affinity has not been lost?
Importantly, there is intrinsic affinity between the CL and CH, plus some additional affinity between the VH and VL, meaning that the overall structure should not change much. Indeed, some people make display libraries of Fabs lacking the disulphide.
thank you for the answers. that is what we thoughts. Now we used TCEP to break the interchain disulfide. How do we know during the Biacore, the disulfide bond will not snap back? Meanwhile how do we know the TCEP does not affect the intrachain disulfide bonds? Any references recommended?
thanks.
there is not selection among all the disulfide bonds based on my experiences.
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