Cancer-specific affinity molecules for diagnosis and targeted therapy
I did MS and PhD from Lucknow University in India and PDF at the University of Tennessee, Memphis. I served (1981-2001) as Scientist A, Scientist B, Senior Scientist C, Project Leader Scientist EI and Principal Scientist EII at Indian Institute of Toxicology Research (a National Lab), Lucknow. I was also a Visiting Scientist (2-year term each) at Ottawa University in Canada, University of Michigan, Ann Arbor, and here at UVM. In 2001, I joined the faculty of the Department of Surgery at UVM.
Our major interest is to develop targeted therapy against cancer. Targeted cancer therapies use drugs that block the growth and spread of cancer by interfering with specific molecules involved in carcinogenesis and tumor growth. By focusing on molecular and cellular changes that are specific to cancer, targeted cancer therapies are believed to be 'magic bullets' that specifically kill cancerous cells without harming to normal cells. We have been interested in identifying molecules that can bind selectively to cancer cells. These affinity molecules are being screened for their effects on cell proliferation, apoptosis, invasion and migration, etc. In combination with toxic chemicals, immunotherapeutic modulators or imaging agents, these affinity molecules can also be used for a selective delivery of required chemicals to cancer cells for a successful killing or diagnostic imaging of tumors. Our research work involves in vitro (fresh tumors, cancer cell lines, purified cancer targets) and in vivo (animals and cancer patients) selection of tumor targeting agents using phage-displayed peptide, single chain antibody and beta-lactamase loop libraries. We are currently focused on generating cancer-specific full-length human antibodies, using EBV immortalization and hybridoma techniques, from B cells obtained from lymph nodes, bone marrow aspirates and tumor tissues of cancer patients. It is hypothesized that utilization of B cells derived from a patient’s immune system that has a pre-existing relationship with a cancer will yield higher quality antibodies with improved specificity to the tumor. The auto-antibodies found to specifically bind tumor cells will be evaluated for their molecular target identification and bioactivity. The long-term objective is to establish methods that reliably and rapidly generate tumor-binding affinity molecules to individual patient tumors which will have broad applicability and provide a platform for developing innovative therapeutic and diagnostic reagents.
Randomized loops of β-lactamase. (A) Schematic representation of different loops (solid black) of the Enterobacter cloacae P99 cephalosporinase molecule, based on the X-ray crystallographic structure, that were selected for randomization. (B) The Molscript drawing of surface structure of the E cloacae P99 cephalosporinase molecule showing that all the selected loops (black) are on one face of the outer surface (based on PDB, 1BLS).
The linear dodecapeptide (X12) and cysteine-constrained decapeptide (CX10C) libraries were created at the N-terminal position of Enterobacter cloacae P99 cephalosporinase (BLA) molecule, between the signal peptide and enzyme protein. The magnified insert region given at the lower left of the figure is inversed in relation to the vector because the insert oligonucleotide and the gene sequences are written 5′– 3′.
Phage particle displaying β-lactamase-random peptide as an N-terminal fusion protein to phage coat pIII. The magnified β-lactamase fusion protein showing a random peptide library at the N-terminal end and FLAG and 6xHis tags at the C-terminal end.
Girja S. Shukla and David N. Krag (2009) Developing bifunctional β-lactamase molecules with built-in target-recognizing module for prodrug therapy: Identification of Enterobacter cloacae P99 cephalosporinase loops suitable for randomization and phage-display selection. Journal of Molecular Recognition, 9 (6): 425-436.
Girja S. Shukla (2009) Phage Display. In: Encyclopedia of Cancer, second edition (ISBN 978-3-540-47648-1), Edited by Manfred Schwab, Springer Press, Heidelberg, Germany.
Girja S. Shukla and David N. Krag (2010) Phage-displayed combinatorial peptide libraries in fusion to β-lactamase as reporter for an accelerated clone screening: Potential uses of selected enzyme-linked affinity reagents in downstream applications. Combinatorial Chemistry and High Throughput Screening, 13 (1): 75-87.
Girja S. Shukla and David N. Krag (2010) Novel β-lactamase-random peptide fusion libraries for phage display selection of cancer cell-targeting agents suitable for enzyme prodrug therapy. Journal of Drug Targeting, 18 (2): 115-124.
Girja S. Shukla and David N. Krag (2010) Cancer cell-specific internalizing ligands from phage displayed β-lactamase-peptide fusion libraries. Protein Engineering, Design & Selection, 23 (6): 431-40.
* indicates equal contribution
CSIR Young Scientist Award The Council of Scientific & Industrial Research (CSIR) introduced, in 1987, a scheme of Awards for Young Scientists in CSIR system in order to promote excellence in various fields of science and technology. These Awards are known as “CSIR Young Scientist Awards”. (Awarded in 1989)
Department of Surgery
Office: E303A Given
Lab: E309 Given
- 12/10/2013 11:30 AM - 12:30 PM
Dr. Kelly Fimlaid
- 12/17/2013 11:30 AM - 12:30 PM
- 1/28/2014 11:30 AM - 12:30 PM
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