TY - JOUR
T1 - Measurement of protein interaction bioenergetics
T2 - Application to structural variants of anti-sCD4 antibody
AU - Doyle, Michael L.
AU - Brigham-Burke, Michael
AU - Blackburn, Michael N.
AU - Brooks, Ian S.
AU - Smith, Thomas M.
AU - Newman, Roland
AU - Reff, Mitchell
AU - Stafford, Walter F.
AU - Sweet, Raymond W.
AU - Truneh, Alemseged
AU - Hensley, Preston
AU - O'Shannessy, Daniel J.
N1 - Funding Information:
Clare Lynn is gratefully acknowledged for experimental assistance. R.W.W. acknowledges the assistance of Narasimha Sreerama in comparisons of the domains of EPOR and grant support from the NIH (GM22994). The authors gratefully acknowledge Frank DiCapua for constructing Figure 16.
PY - 2000
Y1 - 2000
N2 - This chapter has described a bioenergetic analysis of the interaction of sCD4 with an IgG1, and two IgG4derivatives of an anti-sCD4 MAb. The MAbs have identical VHand VLdomains but differ markedly in their CHand CLdomains, raising the question of whether their antigen-binding chemistries are altered. We find the sCD4-binding kinetics and thermodynamics of the MAbs are indistinguishable, which indicates rigorously that the molecular details of the binding interactions are the same. We also showed the importance of using multiple biophysical methods to define the binding model before the bioenergetics can be appropriately interpreted. Analysis of the binding thermodynamics and kinetics suggests conformational changes that might be coupled to sCD4 binding by these MAbs are small or absent.
AB - This chapter has described a bioenergetic analysis of the interaction of sCD4 with an IgG1, and two IgG4derivatives of an anti-sCD4 MAb. The MAbs have identical VHand VLdomains but differ markedly in their CHand CLdomains, raising the question of whether their antigen-binding chemistries are altered. We find the sCD4-binding kinetics and thermodynamics of the MAbs are indistinguishable, which indicates rigorously that the molecular details of the binding interactions are the same. We also showed the importance of using multiple biophysical methods to define the binding model before the bioenergetics can be appropriately interpreted. Analysis of the binding thermodynamics and kinetics suggests conformational changes that might be coupled to sCD4 binding by these MAbs are small or absent.
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U2 - 10.1016/S0076-6879(00)23368-5
DO - 10.1016/S0076-6879(00)23368-5
M3 - Article
C2 - 10944754
AN - SCOPUS:18244373689
SN - 0076-6879
VL - 323
SP - 207
EP - 230
JO - Methods in enzymology
JF - Methods in enzymology
ER -