This year we have continued to identify novel mAbs in several formats as Fabs, scFvs and eAds against cancer-related proteins. These mAbs were tested for their activity against cancer cells in vitro and some of them were also used as tools for conjugation with nanoliposomes and for development of novel approaches for multiple site-specific conjugations of bioactive molecules to antibody fragments. We have also developed several large libraries of eAds which can be used as a source of binders to various targets. The major accomplishments are summarized below. 1) We have previously proposed to use isolated CH2 domains as scaffolds for construction of libraries containing diverse binders, and identified and characterized a stabilized scaffold containing an additional disulfide bond. This scaffold was further characterized and found that by shortening the N-terminus it could be additionally stabilized. This new scaffold is being explored together with the others for generation of libraries of eAds. A specific binder to the membrane proximal external region of the HIV-1 envelope glycoprotein gp41 was selected and is being characterized. 2) Yeast-displayed libraries of engineered CH2 domains were constructed and are being tested for selection of binders. We are also expressing human neonatal Fc receptor (FcRn) to test binding of some of the engineered CH2 domains with the specific aim to generate nAbs that can bind specifically to a cancer-related protein and simultaneously to the FcRn. 3) Previously, we designed a novel type of eAds library containing CDRs in non-cognate positions where H1 was replaced by a library of human light chain CDR3s (L3s) thus combining three most diversified fragments (L3, H3 and H2) in one VH scaffold. The new library was used for selection of eAds against cancer-related proteins, and the selected binders are being characterized. 5) We have continued to characterize our previously identified human mAbs against DR4 and DR5. We also identified a new mAb against the folate receptor beta. This mAb is highly specific and binds with high avidity to cell-surface associated receptor. Currently it is being tested in a mouse model. 6) We have continued to perform high-throughput sequencing of large portions of the antibody repertoires of humans (the human antibodyome) and analyze large number of sequences. We believe that knowledge of the complete antibodyome will have implications for research, diagnosis, prevention and treatment of cancer. It can help for deeper understanding of the B cell (system) biology and diseases, help develop new diagnostic methods based on individual antibodyomes, help predict individual immune responses to immunization and therapeutics as well as provide information for the design of novel therapeutics. Most implications can not be predicted due to the very nature of an omic science.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Investigator-Initiated Intramural Research Projects (ZIA)
Project #
1ZIABC010701-05
Application #
8157402
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
5
Fiscal Year
2010
Total Cost
$588,464
Indirect Cost
Name
National Cancer Institute Division of Basic Sciences
Department
Type
DUNS #
City
State
Country
Zip Code
Ying, Tianlei; Wen, Yumei; Dimitrov, Dimiter S (2017) Precision immunomedicine. Emerg Microbes Infect 6:e25
Li, Dezhi; Gong, Rui; Zheng, Jun et al. (2017) Engineered antibody CH2 domains binding to nucleolin: Isolation, characterization and improvement of aggregation. Biochem Biophys Res Commun 485:446-453
Feng, Yang; Wang, Yanping; Zhu, Zhongyu et al. (2016) Differential killing of CD56-expressing cells by drug-conjugated human antibodies targeting membrane-distal and membrane-proximal non-overlapping epitopes. MAbs 8:799-810
Lynn, R C; Feng, Y; Schutsky, K et al. (2016) High-affinity FR?-specific CAR T cells eradicate AML and normal myeloid lineage without HSC toxicity. Leukemia 30:1355-64
Cherkassky, Leonid; Morello, Aurore; Villena-Vargas, Jonathan et al. (2016) Human CAR T cells with cell-intrinsic PD-1 checkpoint blockade resist tumor-mediated inhibition. J Clin Invest 126:3130-44
Li, Wei; Yang, Hongjia; Dimitrov, Dimiter S (2016) Identification of high-affinity anti-CD16A allotype-independent human antibody domains. Exp Mol Pathol 101:281-289
Lynn, Rachel C; Poussin, Mathilde; Kalota, Anna et al. (2015) Targeting of folate receptor ? on acute myeloid leukemia blasts with chimeric antigen receptor-expressing T cells. Blood 125:3466-76
Ying, Tianlei; Wang, Yanping; Feng, Yang et al. (2015) Engineered antibody domains with significantly increased transcytosis and half-life in macaques mediated by FcRn. MAbs 7:922-30
Chen, Weizao; Li, Wei; Ying, Tianlei et al. (2015) Germlining of the HIV-1 broadly neutralizing antibody domain m36. Antiviral Res 116:62-6
Zhao, Qi; Tran, Hoa; Dimitrov, Dimiter S et al. (2015) A dual-specific anti-IGF-1/IGF-2 human monoclonal antibody alone and in combination with temsirolimus for therapy of neuroblastoma. Int J Cancer 137:2243-52

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