Our current and future research utilizes antibody technology for the generation of a large naive human Fab library in a newly designed phage display vector. This phage display library has become an important source for the generation of human monoclonal antibodies (mAbs) to targets of relevance for hematologic malignancies. We previously selected human mAb KYK-1.0 to human NKG2D from this library. NKG2D is expressed on NK cells which mediate the perhaps most important activity mechanism of antibodies, i.e. antibody-dependent cellular cytotoxicity (ADCC). We further evolved KYK-1.0 through sequential affinity maturation based on phage display technology. The matured antibody (termed KYK-2.0) now binds human NKG2D with subnanomolar affinity. We have been evaluating KYK-2.0 (i) as diagnostic and therapeutic targeting device for NK-cell and T-cell lymphoma and leukemia, (ii) for the generation of bispecific antibodies designed to recruit NKG2D+ NK cells and NKG2D+ CD8+ T cells to the tumor site, and (iii) for interfering with autoimmune processes mediated by NK-cells and T-cells. Complementing our efforts for generating human monoclonal antibodies, we are continuing the mining of rabbit antibody repertoires by phage display. A target of interest in this regard is receptor tyrosine kinase ROR1 which we have found to be uniquely expressed on the surface of B-CLL cells. Using classical hybridoma technology and phage display, we generated a series of mouse and rabbit monoclonal antibodies that bind to different epitopes of ROR1. The characterization of these monoclonal antibodies as well as their utilization for the development of potent antibody-drug conjugates, immunotoxins, chimeric antigen receptors, and bispecific antibodies (which involve above mentioned NK-cell recruiting monoclonal antibody KYK-2.0) is in progress. Another aspect of our antibody engineering efforts is the development of new antibody conjugation technologies. In this area we are closely collaborating with chemists from the Laboratory of Medicinal Chemistry, CCR, NCI, NIH in Frederick, MD. In particular, we are exploring new technologies at the interface of monoclonal antibodies and small synthetic molecules, resulting in uniquely defined immunoconjugates in which the biological and chemical components are endowed with pharmacological advantages. Our focus has been on the 21st natural amino acid selenocysteine as interface between biological and chemical components.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Investigator-Initiated Intramural Research Projects (ZIA)
Project #
1ZIABC010647-07
Application #
8349084
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
7
Fiscal Year
2011
Total Cost
$524,636
Indirect Cost
Name
National Cancer Institute Division of Basic Sciences
Department
Type
DUNS #
City
State
Country
Zip Code
Perez-Galan, Patricia; Mora-Jensen, Helena; Weniger, Marc A et al. (2011) Bortezomib resistance in mantle cell lymphoma is associated with plasmacytic differentiation. Blood 117:542-52
Rader, Christoph (2011) DARTs take aim at BiTEs. Blood 117:4403-4
Ho, Mitchell; Feng, Mingqian; Fisher, Robert J et al. (2011) A novel high-affinity human monoclonal antibody to mesothelin. Int J Cancer 128:2020-30
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Beerli, Roger R; Rader, Christoph (2010) Mining human antibody repertoires. MAbs 2:
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Robak, Laurie A; Venkatesh, Karthik; Lee, Hakjoo et al. (2009) Molecular basis of the interactions of the Nogo-66 receptor and its homolog NgR2 with myelin-associated glycoprotein: development of NgROMNI-Fc, a novel antagonist of CNS myelin inhibition. J Neurosci 29:5768-83
Rader, Christoph (2009) Generation and selection of rabbit antibody libraries by phage display. Methods Mol Biol 525:101-28, xiv
Hofer, Thomas; Skeffington, Lauren R; Chapman, Colby M et al. (2009) Molecularly defined antibody conjugation through a selenocysteine interface. Biochemistry 48:12047-57
Kwong, Ka Yin; Rader, Christoph (2009) E. coli expression and purification of Fab antibody fragments. Curr Protoc Protein Sci Chapter 6:Unit 6.10

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