Antibody Name: anti-B4
References: Roguska, M.A., Pedersen, J.T., Henry, A.H., Searle, S.M.J., Roja, D.M., Avery, B., Hoffee, M., Cook, S., Lambert, J.M., Blattler, W.A., Rees, A.R. and Guild, B.C. "A comparison of two murine monoclonal antibodies humanized by CDR-grafting and variable domain resurfacing" Prot. Engng. (1996) 9:895-904.
Roguska, M.A., Pedersen, J.T., Keddy, C.A., Henry, A.H., Searle, S.J., Lamber, J.M., Goldmacher, V.S., Blattler, W.A., Rees, A.R. and Guild, B.C. "Humanization of murine monoclonal antibodies through variable domain resurfacing" Proc. Natl. Acad. Sci. (1994) 91:969-973.
(see also mAb N901).
Donor Antibody: mouse
Acceptor Antibody: Human IgG1 | k
Antigen: CD19 antigen on human B cells
Laboratory: ImmunoGen, MA; University Bath, UK
Design: Models of mouse and humanised V regions generated by AbM. Analyse FR residues within 6A of a CDR for changes in size, charge, hydrophobicity or potential to form H-bonds that could disturb the CDR conformation.
Frameworks: Two approaches:
Highest identity VH 21/28 | VL POP
Clonally derived pair VH LS5 | VL LS5
CDRs: All 6 VH and VL, Kabat Definition except that CDR-H2 was 7 residues shorter at the C-terminus (note however: backmutations 60-64 revert CDR-H2 to Kabat Definition)
Backmutations: Highest identity:
VH R44G S60N Q64K R71V T73K
VL L46R L47W D70S F71Y T72S V104L G108R
Clonally derived:
VH A60N N62K L63F Q64K T71V T73K
VL L46R D70S F71Y
Binding: Clonally derived gave 1/3 binding to Namalwa cells as did mouse mAb. Highest identity FRs gave binding "equal" to that of mouse mAb.
Expression: transient expression in cos cells
Comment: This mAb was also resurfaced.
Clinical Indication: B cell malignancies

explanation of symbols and abbreviations

José Saldanha © 1997-8. Birkbeck College, London WC1E 7HX.