Monoclonal antibody lead characterization: in vitro and in vivo methods.
This chapter describes in vitro and in vivo methods to characterize a lead monoclonal antibody candidate in the drug discovery setting. Approaches to characterize monoclonal antibody specificity, heavy and light chain composition, and antibody mode of action including the ability to mediate secretion of effector molecules, inhibit cell proliferation, induce apoptosis, or elicit antibody effector function are described. ELISA and flow cytometry based methods, as well as in vitro assays to assess for cell proliferation, ADCC, and CDC are detailed.In addition, both subcutaneous and orthotopic in vivo tumor xenograft models to assess antibody efficacy are described. The xenograft tumor model is a valuable tool for assessing the therapeutic activity of a monoclonal antibody drug candidate. Xenograft models are generated by the implantation of tumor cells or tumor fragments of human origin into immune-compromised mice or rats. This allows for fast and efficient in vivo evaluation of an antibody drug candidate in human cancer models. Here, we describe the procedures for generating preclinical animal tumor models frequently employed in the preclinical drug discovery setting.
AstraZeneca R&D Boston, Waltham, MA, USA.
Axel
A
Hernandez
Julie
J
Parmentier
Youzhen
Y
Wang
Jane
J
Cheng
Gadi Gazit
GG
Bornstein
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000920:Antibody-Dependent Cell Cytotoxicity / Q000276:immunology; D017209:Apoptosis; D049109:Cell Proliferation; D003165:Complement System Proteins; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D008297:Male; D051379:Mice; D008967:Molecular Biology / Q000379:methods*; D051381:Rats; D023041:Xenograft Model Antitumor Assays
D000911:Antibodies, Monoclonal; D003165:Complement System Proteins