TCR/CAR discovery

Rukmini Ladi

*** ******** ****, ***** *********, MA 01862* 317-***-**** * ***********@*****.***

EXECUTIVE SUMMARY

• Research Scientist with more than 7 years’ experience in immuno-oncology, gene and cell therapy, drug discovery and formulation development

• Extensive experience with assay development, optimization and multi-color flow cytometry (FACS)

• Hands on experience in cell/molecular biology, gene editing techniques, immune based in-vitro assays on primary human T-cells and iNKT cells, in-vivo mouse studies and ex-vivo mouse tissue analysis

• Strategic and collaborative team player with ability to multi-task and successfully achieve multiple project goals

• Demonstrated expertise in scientific project management enabling high quality and on time results KEY FUNCTIONAL AND LEADERSHIP SKILLS

Functional Skills Leadership Skills

• Adoptive immunotherapy

(TCR/CAR in T-cells and iNKT cells)

• Immunoassays and cell culture

• NextGen sequencing/single cell analysis

• Formulation development and characterization

• Innovation

• Strategic Thinking

• Relationship Building

• Influence

• Collaboration

RESEARCH SKILLS

Cell Biology and Immunology (CAR-T/T-cell Therapy): Isolation and expansion of primary human T-cells, B-cells and iNKT cells, human PBMC separation, immunophenotyping, multi-color flow cytometry (14+ colors, BD Fortessa/BD Canto), T-cell and iNKT cell transduction and transfection

(electroporation), immune cell co-culture/functional assays, cytotoxicity assays, multi-plex cytokine release assays

(Luminex platform), cell culture: suspension/adherent mammalian and human cell lines, cellular imaging Molecular Biology:

In-vitro RNA synthesis, RT-PCR/qPCR, SDS-PAGE, ELISA, western blotting, cloning, plasmid extraction and purification, ligation, transformation, siRNA design and gene silencing, isolation and purification of DNA and RNA from tissues and cells, NextGen sequencing and analysis of human B-cells and T-cells, virus quantification, virus neutralization assay, antibody purification, gel extraction and purification, electrophoretic mobility shift assay, BCA, MTT assay Analytical Instruments and Techniques:

DSC, HPLC, NMR, UV/Vis Spectrophotometry for nucleic acid quantification, Dynamic Light Scattering, thin layer/column chromatography, USP dissolution testing apparatus and disintegration RESEARCH EXPERIENCE

Senior Research Associate

AgenTus Therapeutics, Lexington, MA, January 2020-April 2020

• Independently led the process development of the allogeneic adoptive cell therapy project to enhance recombinant TCR expression in iNKT cells by optimizing lentivirus transduction protocols

• Successfully designed, optimized and established protocol for transfections in iNKT cells via siRNA delivery method

• Lead process development of iNKT cell culture, stimulation and in-vitro expansion using B-cells from PBMCs

• Designed and executed in-vivo studies using xenograft mouse models

• Mentored Northeastern University co-op student on RNAi research project and trained on technical research skills- flow- cytometry, qRT-PCR, RNA/DNA isolation and purification, immunological assays and data analysis Research Associate

AgenTus Therapeutics, Lexington, MA, December 2018-December 2019

• Technical lead on autologous T-cell immunotherapy project using receptor discovery platform to screen novel cancer- specific T-cell receptors (TCRs)

• Engineered antigen-specific primary T-cells and iNKT cells by lentiviral transductions and mRNA transfections with CAR-T and TCRs

• Designed and executed various in-vitro immunological assays such as luciferase and flow cytometry-based cytotoxicity assays, multiplex cytokine and chemokine assays and immune cell co-culture assays to validate anti-cancer TCRs

• Identified, characterized and optimized the CAR-T and TCRs by immunophenotyping with multiparameter flow cytometry and by performing quantitative real-time PCR (qRT-PCR)

• Designed, processed and analyzed engineered human primary T-cells from in-vivo tissue samples for gene expression and vector copy number (VCN) using qRT-PCR. Data generated was used in CTA filing for the NYESO-1 TCR program.

• Optimized and established non-viral transfection protocol to generate in-vitro transcribed RNA (IVT) for rapid TCR engineering of T-cells and iNKT cells

• Trained co-op student’ on technical laboratory skills such as aseptic cell culturing (tumor/lymphocytes), qRT-PCR, and flow cytometry

• Meticulously maintained electronic lab notebook and documented experimental findings

• Effectively presented research findings to senior leadership and colleagues Graduate Research Assistant

University of Kansas, Lawrence, KS, July 2016-August 2018

• Cell line development: Prepared single-cell mammalian and human cell cultures under stimulated and unstimulated conditions to evaluate the expression of receptors using in-vitro models

• Human immune cell isolation: Co-ordinated and mentored PBMC isolation for B-cell/T-cell receptor analyses

• Established and (cryo)preserved in-vitro human B-cell and T-cell stimulation cultures for NextGen sequence analyses and functional cell-based assays

• Performed FACS single cell sorting to identify anti-cancer functional BCR and TCR repertoire sequences

• Directed experiments and data analysis that led to discovery of new experimental pipeline for NextGen sequencing natively paired human T-cell receptor repertoires from single lymphocyte cells

• Initiated and developed anti-cancer BCR/TCR libraries by cloning into lentiviral expression vector

• Troubleshooted, planned and managed multiple research projects on exploring, investigating and analyzing antibody repertoire sequences from EBV, PTLD and healthy patients

• Facilitated virus quantification and neutralization assays by adapting innovative experimental protocol

• Organized and inventoried laboratory merchandise and office supplies to increase the accuracy and speed of interlaboratory orders

Graduate Research Assistant

Butler University, Indianapolis, IN, August 2013-May 2016

• Formulated siRNA complexes with modified β cyclodextrin nanoparticles and liposomes for mutant huntingtin gene knockdown in PC-12 cell line

• Synthesized cationic β cyclodextrin nanoparticles by one-step condensation polymerization method to form an electrostatic complex with siRNA

• Led the production of mutant huntingtin protein in PC-12 cell line as an in-vitro model for Huntington’s disease

• Characterized formulation for optimum particle size, surface charge, practical yield, siRNA complex formation, monomeric substitution, phase transition behavior, cell cytotoxicity and gene knockdown

• Analyzed and documented data using Spin Works 4, OriginLab Graphing and Analyzing software Quality Control Chemist

Serum Institute of India, Pune, India, July 2012–December 2012

• Utilized laboratory instruments such as pH meter, UV/Vis spectrophotometry, liquid particle counter, Atomic Absorption Spectrophotometer and Capillary zone electrophoresis for analyzing batch-to-batch uniformity of vaccine vials

• Evaluated data and prepared reports for component specifications in compliance with cGMP, cGLP and FDA by using Microsoft Excel spreadsheet

• Ensured batch uniformity and integrity by communicating and submitting analysis reports to the manufacturing department

EDUCATION

Master of Science in Pharmaceutical Chemistry/Immune Engineering University of Kansas, Lawrence, KS, 2018

Master of Science in Pharmaceutical Sciences (Pharmaceutics) Butler University, Indianapolis, IN, 2016

Bachelor of Pharmacy

Pune University, Pune, India, 2012

ACADEMIC HONORS

• Recipient of Graduate Research Excellence Award by Butler University; 2016

• Best Outgoing Student Award by Pune University; 2012

• Top 10 Student Recognition Award by Pune University, 2012

• Ranked 1st in academics throughout the 4 years of Bachelor of Pharmacy course; 2008-2012 LICENSURES

Licensed Pharmacist in the state of Maharashtra, India, September 2012–Present License number: 134527

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