A humanized aldolase antibody for selective chemotherapy and adaptor immunotherapy.
Mouse monoclonal antibody 38C2 is the prototype of a new class of catalytic antibodies that were generated by reactive immunization. Through a reactive lysine, 38C2 catalyzes aldol and retro-aldol reactions using the enamine mechanism of natural aldolases. In addition to its remarkable versatility and efficacy in synthetic organic chemistry, 38C2 has been used for the selective activation of prodrugs in vitro and in vivo and thereby emerged as a promising tool for selective chemotherapy. Adding another application with relevance for cancer therapy, designated adaptor immunotherapy, we have recently shown that 38C2 can be chemically programmed to target tumors by formation of a covalent bond of defined stoichiometry with a beta-diketone derivative of an integrin alpha(v)beta(3) targeting RGD peptidomimetic. However, a major limitation for the transition from preclinical to clinical evaluation is the human anti-mouse antibody immune response that mouse 38C2 is likely to elicit in a majority of patients after single administration. Here, we report the humanization of mouse 38C2 based on rational design guided by molecular modeling. In essence, the catalytic center of mouse 38C2, which encompasses a deep hydrophobic pocket with a reactive lysine residue at the bottom, was grafted into a human antibody framework. Humanized 38C2 IgG1 was found to bind to beta-diketone haptens with conserved affinities and revealed strong catalytic activity with identical k(cat) and slightly higher K(M) values compared to the parental mouse antibody. Furthermore, humanized 38C2 IgG1 revealed efficiency in prodrug activation and chemical programming comparable to the parental mouse antibody.
Rader
Christoph
C
The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037, USA. raderc@mail.nih.gov
Turner
James M
JM
Heine
Andreas
A
Shabat
Doron
D
Sinha
Subhash C
SC
Wilson
Ian A
IA
Lerner
Richard A
RA
Barbas
Carlos F
CF
D000595:Amino Acid Sequence; D000818:Animals; D017151:Antibodies, Catalytic / Q000737:chemistry / Q000627:therapeutic use; D020134:Catalytic Domain; D016503:Drug Delivery Systems; D005634:Fructose-Bisphosphate Aldolase / Q000737:chemistry / Q000627:therapeutic use*; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry / Q000627:therapeutic use*; D007167:Immunotherapy; D007659:Ketones / Q000737:chemistry / Q000378:metabolism; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D015394:Molecular Structure; D011355:Prodrugs / Q000378:metabolism; D015202:Protein Engineering; D016415:Sequence Alignment
D017151:Antibodies, Catalytic; D007140:Immunoglobulin Fab Fragments; D007659:Ketones; D011355:Prodrugs; C109536:antibody aldolase; D005634:Fructose-Bisphosphate Aldolase