Stability of murine, chimeric and humanized antibodies against pre-S2 surface antigen of hepatitis B virus.
We have constructed a humanized antibody with specificity for the pre-S2 surface antigen of hepatitis B virus (HBV) by grafting the complementarity determining regions (CDRs) of parental murine monoclonal antibody (mAb) into human anti-Sm antibody framework regions. The humanized antibody has a substitution at position 94 in a framework region of the heavy chain variable region, and exhibits the same antigen binding affinity as the parental murine monoclonal and chimeric antibodies. In order to assess the stability of these antibodies, thermal inactivation of the parental, chimeric and humanized antibodies was analyzed. Fifty percent inactivation of the chimeric and humanized antibodies was observed at 63.7 degrees C and 68.7 degrees C, respectively, compared to 55.0 degrees C for murine antibody. The humanized antibody also exhibited increased stability against denaturant. Guanidine-induced unfolding monitored by the changes in fluorescence intensity at 360 nm showed that midpoints of the transition of the chimeric and humanized antibodies were 2.47 M and 2.56 M, respectively, whereas that of the murine antibody was 1.36 M.
Park
Sung Sup
SS
Korea Research Institute of Bioscience and Biotechnology (KRIBB), 52 Oun-dong, Yusong, 305-333, Taejon, South Korea. sspark@kribb.re.kr
Kim
Jeongho
J
Brandts
John F
JF
Hong
Hyo Jeong
HJ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D022801:Complementarity Determining Regions; D004355:Drug Stability; D019791:Guanidine; D006510:Hepatitis B Antibodies / Q000737:chemistry; D006514:Hepatitis B Surface Antigens / Q000276:immunology*; D006801:Humans; D051379:Mice; D011489:Protein Denaturation; D011498:Protein Precursors / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000276:immunology; D013816:Thermodynamics; D014759:Viral Envelope Proteins / Q000276:immunology*
D000911:Antibodies, Monoclonal; D022801:Complementarity Determining Regions; D006510:Hepatitis B Antibodies; D006514:Hepatitis B Surface Antigens; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; C050807:presurface protein 2, hepatitis B surface antigen; D019791:Guanidine