Summary:

• Manage timelines for supporting product testing and characterization to meet organizational objectives
• Align project strategies and priorities in managing complexities for business needs
• Work effectively in cross-functional teams including Quality unit, R&D, Formulation and Production team in a coordination effort for accomplishing projects objectives

Professional Experience:

Confidential, Old Saw Mill River Road, Tarrytown, NY  
Scientist 12/2006-06/2011; Sr. Scientist QC/AD 07/2011-11/2011

• Responsible for new analytical method development leading to development, optimization, transfer and validation of methods in protein chemistry and antibody chemistry, find out and addressed root cause of investigations, developed associated protocols, procedures and reports for documentation as prioritized.
• Managed and applied in-depth knowledge of HPLC, such as Ion Exchange (IE), Normal Phase (NP), Reversed Phase (RP), SEC with either UV (DAD), FLD, RI, or ELSD detection for establishing conformational and structure function relationship of antibodies. Accomplished projects activities and performed work in compliance with cGMP, safety and regulatory ICH/FDA guidelines.
• Coordinated Director of Analytical Development, and Supported effectively for QC routine analytical activities such as: release and characterization testing, monitoring stability for stability batches, raw materials and in-process materials testing, as well as completing documentation in support to meet timeline.
• Performed stability and release testing of products to conform specifications, and tested for physico-chemical and conformational characterization, product-related impurities, as well as degradation pathway studies of clinical trial lots.
• Participated clinical and preclinical testing of biologics (Reference standard, drug substance and drug product), and comparability studies of new products purified either at PGNX or at CMO/CRO and delivered test results for supporting strict FDA timeline.
• Provided cross-functional analytical support for new product development, and supported analytical activities for excipients assay development and validation, or trouble-shooting activities for manufacturing, quality control and formulation development.
• Responsible for QC/AD Instruments service and maintenance. Coordinated vendors for executing scheduled IQ/OQ/PQ of Agilent Chemstation/Chemstore HPLC Systems, Performed PQ, and evaluated systems for normal operating conditions with installed ChemStation Plus Security Pack software 21CFR Part 11.
• Presented progress report and strategies in QC/AD department meeting and associated meetings for project management activities within Quality unit, R&D department, Formulation department and Production Teams in coordination effort for accomplishments objectives and for supporting Analytical characterization.
• Authored SOPs, investigation reports, protocols and summary reports, method development and validation reports, stability and batch studies documents. Wrote and Reviewed QC test documents, Change Control and Laboratory notebooks prior to submission for QA approval. Prepared stability data tables for supporting Regulatory CMC.
• Provided Training to QC/AD Scientists on various procedures followed through SOPs, Compliances, as well as guided for new method development and resolving Instrumental or technical issues as assigned. Co-ordinated team for proper maintenance and operations in the laboratory cGMP.

List of the analytical methods developed, Validated and Transferred to QC. SOPs include but not limited to:

• SOP_Quantitative Determination of Histidine in PRO 140 (CHO) 10-180 mg/mL Bulk Drug Substance and Finished Drug Product Using Reversed Phase High Performance Liquid Chromatography and UV Detection
• SOP_Quantitative Determination of Sucrose in PRO 140 (CHO) 10-180 mg/mL Bulk Drug Substance and Finished Drug Product Using Normal Phase High Performance Liquid Chromatography and Refractive Index Detection (Agilent 1100/1200)
• SOP_Quantitative Determination of Glycine and Histidine in PRO 140 (CHO) Finished Drug Product 175mg/mL (Histidine/Glycine/Sorbitol/Sodium Chloride/Polysorbate 20 Buffer)
• SOP_RP-HPLC Tryptic Peptide Mapping of PRO 140 (CHO) 10-180 mg/mL Bulk Drug Substance and Finished Drug Product
• SOP_RP-HPLC Tryptic Peptide Mapping of PSMA mAb and PSMA ADC FDP
• SOP_Determination of Deamidation Levels in PSMA, PSMA Drug Conjugate (PSMA ADC) and PRO 140 (10-180mg/mL) Monoclonal Antibody by Isoquant Isoaspartate Detection Kit Followed by Strong-Cation Exchange HPLC Method
• SOP_Quantitative Determination of Polysorbate 20 in PSMA ADC Finished Drug Product Using Reversed Phase High Performance Liquid Chromatography and ELSD Detection
• SOP_Identification of Tryptophan in CD CHO Feed Concentrate Part E Raw Material
• SOP_Identification of Glucose in CD CHO Feed Concentrate Part C (50X) Raw Material
• SOP_Identification of Glucose in HyClone Cell Boost 1TM (R05.2) Raw Material
• SOP_Determination of Non-Proteinaceous Impurities by Reversed Phase HPLC (The method was originally developed at Piramal. Trouble shoot, validated and introduced to QC testing). Validation Protocol and Validation Report for the Quantitation of Non-Proteinaceous Impurities in PSMA ADC Finished Drug Product

List of the analytical methods developed for mAb and ADC characterization include but are not limited to:

• DTM_Quantitative Determination of Instant labeled N-glycans of PSMA ADC FDP and PSMA ADC In-process Samples using NP-HPLC
• DTM_Quantitative Determination of Glycine and Histidine in PSMA ADC Finished Drug Product using mixed-mode Reversed- Phase and Cation-Exchange HPLC
• DTM_Quantitative Determination of Residual Dimethyl Sulfoxide (DMSO) Content in PSMA ADC BDS and FDP
• DTM_ SEC-HPLC Analysis of reduced Prostate Specific Membrane Antigen Monoclonal Antibody (PSMA mAb) and reduced Prostate Specific Membrane Antigen Monoclonal Antibody Drug Conjugate (PSMA ADC). The light chain fractions and heavy chain fractions were collected, purified and lyophilized for MS characterization
• DTM_Detection of Subvisible Particles in monoclonal antibodies formulations using BD FACSCalibur

Training to intervene regulatory compliances:

• Attended Refresher’s training on GMP, and Corporate Governance training
• Attended in-house potent compound handling training for improved understanding on handling of potent compounds
• Attended training for Record Management System
• Attended in-house Annual Health and Safety Awareness Training
• Attended free webinars, training on company policies and regulations
• Attended training and Explored HPLC operation using an updated version of Chemstation, Chemstore and Security Pack for 21 CFR Part 11 compliance and electronic signatures

Confidential, American Road, Morris Plains, NJ 
Scientist, Analytical R&D 1/2002-11/2005
Development, Qualifications of analytical methods for characterization and lot release comparability of monoclonal antibodies followed:

• Developed a normal phase HPLC method for quantitative determination of oligosaccharides in humanized monoclonal antibodies with fluorescence detection under Waters HPLC. The NP-HPLC Glycan analysis method was Qualified. Authored method development SOP and the method Qualification report
• Developed a reversed phase HPLC method for quantitative determination of monosaccharides in humanized monoclonal antibodies with fluorescence detection under Waters HPLC and authored the test method SOP.
• The Assignments and mapping of N-glycans were accomplished by treating glycans with highly specific exoglycosidase
• Developed quantitative method of cation exchange chromatography, CEX-HPLC, under low pH separation of different sub-forms of C-terminal lysine variants
• Developed reversed phase HPLC peptide mapping method for Qualitative characterization of primary structure of antibodies, Fab and Fc, antibodies conjugates
• Developed the method of LCA HPLC and isolated defucosylated and fucosylated population of monoclonal antibodies formulations for studying monoclonal antibodies effector functions
• Researched optimization of new methods of development, analytics of evaluation impurities, degradants, stability testing of in process and finished products, Lot release comparability tests, Data acquisition and Statistical analysis in accordance with GMP, USP and ICH guidelines
• Managed and responsible of a brand new laboratory suited to the Garden state Cancer Center, setting up the laboratory for Yeast and bacterial fermentation in BioFloR 10L bench top fermenter under cGMP
• Build up the fermentation production system, Developed cell lines of E.coli and P. pastoris, developed high throughput industry methods including Fed-batch, batch, continuous fermentation pilot scale production processes, and downstream purification procedures for divalent, trivalent, bispecific fusion proteins engineered from antibody variable domains, manufacturing of Industry proteins, purification and characterization .
• Designed and performed experimental methodologies, led the team sharing workings
• Interacted with in house Quality Control laboratory, Chemistry laboratory, and production facility to perform analytical characterization of in-process and finished pharmaceuticals, and antibody drug conjugates
• Supported Regulatory managements by testing lot release samples, providing and supplementing lot release comparability data for NDA submission to approval
• Reviewed FDA NDA/INDA outcomes and applied the instructions as needed
• Wrote technical reports, SOP, Summary, batch records of all processes
• Attended, participated, and presented in managerial meetings, group meetings, seminars
• Transferred HPLC test methods and qualified procedures to QA, QC and Regulatory managements

Confidential, Kingsland street, Nutley, NJ
Biochemist, R&D 6/2000-7/2001
Responsible for HTS method development of B. subtilis and E. coli GeneChip microarray, coordinated Quality control assessment, developed new overproducer strains for high scale biotin production, Supported the Team achieving goal and meeting timelines.

• Developed RNA expression analysis using an antisense B. subtilis genome Array
• Isolated biotin-labeled cDNA targets from shaker flasks or fermentation cell culture of Vitamin overproducer
• Isolated and purified total RNA using Macaloid Clay/SDS beads extraction
• Biotin labeled cDNA targets, Hybridization with either B.subtilis Sense or E.coli antisense GeneChip, Detection under Confocal laser microscope and Digital Image Data file, GeneSpring Data analysis, Statistical data analysis/Data mining
• Analyzed biotin-regulated gene expression, Clustering analysis, Functional assignment IntraNet Annotated Expression Database (Subtilis/Micado/Sub2D)
• Tested GeneChip Quality Control for Temperature, Dynamic range cDNA hybridization, Differential gene expression for biotin, Riboflavin, thiamin treatment
• Several gene clusters identified in B. subtilis GeneChip expression were regulated by biotin
• Constructed Non-polar insertion and deletion mutant strains to evaluate biotin overproduction in high scale fermentations
• Developed HTS RNA expression method using a sense E. coli GeneChip and screened potential strains of E. coli
• By use of both B. subtilis and E. coli GeneChip microarrays, both genomes were exposed with bacimethrin-a naturally occurring thiamin antimetabolite and identified novel gene expression for biotin and thiamin overproduction

Mycology Laboratory, Confidential, Albany, NY
Post doctoral Research Affiliate 9/1998-5/2000
Research objectives aimed to the discovery and development of antifungal drugs: Master the biology and molecular genetics of pathogenic fungi and identified sequences of promising targets for stress response and signal transduction pathway of Cryptococcus neoformans (Cn) and Candida albicans (Ca)

• Reported first that the, GPD1, encoding sn-glycerol-3-phosphate dehydrogenase (GPD), the key enzyme responsible for the biosynthesis of glycerol in Cn
• Cloned and sequenced the GPD1, encoding sn-glycerol-3-phosphate dehydrogenase by screening and complementing S. cerevisiae GPD1Δ mutant (glycerol deficient and salt sensitive) and with a Cn yeast expression library
• A genomic CnGPD1cosmid clone with a 4.5 kb insert was isolated, sequenced and constructed null CnGPD1Δ mutant
• sn-glycerol-3-phosphate dehydrogenase (NAD+) gene of C. albicans was cloned and sequenced.
• Cloned and sequenced the ADH1, encoding alcohol dehydrogenase, a partial transcript and EXO1, encoding exo-ß-glucanase gene of Cn
• Cloned and sequenced the cDNA and the genomic version of HOG1, the mitogen-activated protein kinase gene of Cn and studied pathogenecity in mouse model

Department of Biochemistry, Confidential, NJ, Piscataway, New Jersey 
Postdoctoral Fellow 9/1996-5/1998 
Research Interest and objective aimed on: Elucidation of signal transduction mechanisms underlying regulation of EnvZ kinase and phosphatase activities in E. coli
Analyzed cytoplasmic domain of EnvZ by dividing it into two small sub-domains (A and B) which are able to reconstitute the kinase function when mixed.
Sub-domain A consists of 67 residue forming a stable dimer, which has the autophosphorylation site and the binding region of the downstream response regulator. Sub-domain B consisting of 161 residue exists as a monomer and functions as the kinase.

• This two-domain structure provides for the first time, an insight into the structural arrangement of the enzyme and its enzymatic mechanism.
• Developed optimized protein purification processes and achieved quality products of recombinant proteins (≥ grams level) in E. coli cell culture using shaker flasks labeled with 13C and 15N
• Developed processes for purification to homogeneity, and the purified protein molecules were used for Three dimensional structure determination of both domains by NMR
• The high scale purification procedures allowed a significant reduction (≥1000 fold) of manufacturing cost of quality products and enhanced the three dimensional structure determination of histidine kinase of E.coli.

Interaction of these two sub-domains, as well as NMR structure of sub-domain A bound to OmpR and kinetic characterization of kinase and phosphotransfer function of EnvZ were addressed for a precise demonstration of the mechanism of signal transduction of Histidyl-Aspartyl phosphorelay system in E. col.

Confidential, Dept. of Biochemistry and Mol. Biol. Urawa 338, Japan
Monbusho Research Fellow/doctoral1/1993-3/1996

Worked on Biosynthetic regulations and biological functions of phospholipids in E. coli includes:

Constructed a null pssA mutant of E. coli by replacing the pssA gene with a drug resistance gene
Replaced the soluble phosphatidylserine synthase with membrane bound PS synthase of B. subtilis and roled out the necessity 
of membrane bound form

Discovered the incompatibility of null pssA and cls alleles, and the essentiality of acidic phospholipids (PG and CL) 
for zwitterionic phosphatidylethanolamine (PE) synthesis

Constructed an inactivated allele of B. subtilis phosphatidylserine synthase by using modified primers in PCR amplification on its structural gene, pss and Explored the deleterious effect of overproduced PE in E. coli

A Cross Feedback Model was established which explain the balanced synthesis of zwitterionic and acidic phospholipid in E. coli

Dept. of Microbiology, Confidential, Dhaka, Bangladesh 
Research Fellow/NCST 7/1991-12/1992
Research objective aimed on Characterization of extracellular protease activity of Bacillus thuringensis strains pathogenic to mosquito larvae and development of low cost media for its production as biopesticide
Accomplishments: 
Discovered the Efficacy of different drugs on clinical isolates of Salmonellae, Streptococci, Bacilli, etc. in Bangladesh

TECHNICAL EXPERTISE:

Analytical Assays: RP-HPLC, NP-HPLC, CEX-HPLC, SCX-HPLC, SEC-HPLC, Tryptic Peptide Mapping, Lectin-ConA, LCA HPLC Waters Millenium (UV, FLD), In vitro phospholipids analysis 2D TLC, BD FACSCAN and BD FACSCALIBUR Flow Cytometer, Spectroscopy Shimadzu (UV), Agilent 1200 Chemstation/Chemstore/21CFR part 11 HPLC (UV, FLD, RI, ELSD) 

CHIP based Assays: HTS RNA expression, Antisense/sense B. subtilis , E. coli GeneChip microarray, Affymetrix GeneChip/GeneSpring, Data mining

Drug development processes: Fusion constructions of diabody, triabody, bispecific etc consisting heterologous polypeptide chains from the variable domains of humanized anti-CEA antibody, labetuzumab and h679, binding sites for HSG (histamine-succinyl-glycine) and with 6H 

In vitro glycerol analysis, Determination of MIC and PAE of antibiotics, Fluorescence Microscopy, Gram staining, 

The role of glycerol in fungal pathogenesis were studied in animal model. BALB/c mices were challenged with different doses of the mutant and parental gpd1 cells by inoculation into the lateral vein of the tail, and evaluated pathogenecity.

Protein Chemistry: Stable cell lines of Pichia pastoris, E. coli, B. subtilis development, Purification and characterization of native and recombinant proteins; ion exchange, hydrophobic interaction, Ni-NTA, gel filtration and affinity chromatography using bench-top columns, IMAC, IEC, Size exclusion HPLC (Beckman), FPLC (Pharmacia, LCC-500), SDS-PAGE, Silver stained Native, ELISA, BIAcore, buffer exchange, and filtration processes using ultrafiltration (Millipore/Amicon Stirred cell), diafiltration techniques, TFF/UF membrane filtration, concentration Amicon, membrane protein isolation, purification, production of recombinant proteins in Pichia pastoris, E. coli, B. subtilis using both shaker flasks and pilot fermentors (New Brunswick BioFlo) followed cGMP, Continuous fermentation, Fed-batch fermentation, process optimization, scale-up etc 

Molecular Biology: Cloning and expression in both prokaryotic and eukaryotic systems; DNA & RNA isolation, purification, sequencing, mapping and assays, Northern blot, Southern blot, Dot blot, PCR, Deletion mutant construction, Site-directed mutagenesis, auxotrophic mutant isolation, phage display libraries; colony hybridization, Radiolabeling, Screening of Yeast two hybrid cDNA libraries, Cloning cDNA and genomic DNA, In vitro enzyme assay, Histidine Kinase assay, Phosphatase assay, Phosphatidyl serine synthase assay, engineering of recombinant proteins, DNA and protein database search and sequence analysis, GCG (Genetics Computer Group)

PUBLICATIONS:

• Lee, JM., Zhang, S., Saha, SK., Anna, SS., Jiang, C., and Perkins, J. (2001) RNA expression analysis using an anti-sense Bacillus subtilis genome array,
• J. Bacteriol, 183 (24):7371-7380
• Reddick, JJ., Saha, SK., Lee, JM., Melnick, JS., Perkins, J., and Begley, TP (2001)The mechanism of action of bacimethrin, a naturally occuring thiamin antimetabolite, Bioorganic and Medicinal Chemistry Letters, 11(17): 2245-2248
• Tomomori, C., Tanaka, T., Dutta, R., Park, H., Saha, S.K., Zhu, Yan., Ishima, R., Liu, D., Tong, K.I., Kurokawa, H., Qian, H., Inouye, M., and Ikura, M. (1999) Solution structure of the homodimeric core domain of E. coli histidine kinase EnvZ,
• Nature Structural Biology, 6(8):729-34
• Tanaka. T., Saha. S. K.,Tomomori. C.,Ishima. R, Liu. D.,Tong. K. I., Park. H., Dutta. R., Qin. l., Swindells. B. M.,Yamazaki. T., Ono. M. A., Kainosho. M., Inouye. M, and Ikura. M. (1998) NMR structure of the histidine kinase domain of the E. coli osmosensor EnvZ,
• Nature, 396:88-92
• Park, H., Saha, S. K., and Inouye, M. (1998) Two-domain reconstitution of a functional protein histidine kinase,
• Pro. Nat. Aca. Sci., 95:6728-6732
• Saha, S. K., Nishijima, S., Matsuzaki, H., Shibuya, I., and Matsumoto, K. (1996) A regulatory mechanism for the balanced synthesis of membrane phospholipid synthesis in E. coli., Biosci.Biotech.Biochem., 60:111-116
• Saha, S. K., Furukawa, Y., Matsuzaki, H., Shibuya, I., and Matsumoto, K. (1996) Directed mutagenesis, Ser-56 to Pro, of Bacillus subtilis phosphatidylserine synthase drastically lowers enzymatic activity and relieves amplification toxicity in E. coli, Biosci. Biotech. Biochem., 60:630-633
• Saha, S. K., and Saha, S. K., (1994) Antibiotic resistance of Salmonella typhi in Bangladesh,
• J. Antimicrob. Chemother., 33: 190-191
• Hossain, A., Saha, S.K., Akhtaruzzaman, S., Kabir. Y., and Rahman, M. (1994) Studies on extracellular protease activity from locally isolated Bacillus thuringiensis strains. D.U. J. Biol. Sci. 3: 83-87

ABSTRACTS:

• E. A. Rossi, S. K. Saha, R. M. Shakey, H. Karacay, I.D. Horak, D. M. Goldenberg and C. H. Chang. (2004) A trivalent bispecific fusion protein engineered from antibody variable domains for improved pretargeting of CEA-expressing cancers, AACR 95th Annual Meeting, Orlando, Florida, USA, March 27-31
• Dyavaiah M, Saha SK, Chaturvedi S & Chaturvedi V (2005) Glycerol is the major polyol of Candida albicans and Cryptococcus neoformans with important roles as stress protectant and antioxidant. Abstr. F-165, 106th General Meeting, Amercian Society for Microbiology, Orlando, FL.
• Saha, S. K., and Chaturvedi, V. (1999) Characterization of sn-glycerol-3-phosphate dehydrogenase (NAD+) gene of Cryptococcus neoformans:Role in stress response and virulence. Interscience Conference on Antimicrobial Agents and Chemotherapy (ICAAC), San Francisco, USA, Sep 26-29, 1999
• Saha, S.K., Okada, M., Matsuzaki, H., Shibuya, I., and Matsumoto, K. (1994) Biological roles of phospholipids: an Escherichia coli mutant completely lacking the structural gene for phosphatidylserine synthase. Abstract of the 67th annual meeting of Japanese Biochemical Society, Osaka, Japan, Seikagaku 66:800

AWARDS/Scholarship:

• Monbusho Fellowship, By Japan Government for research studentship and doctoral program (Jan 1993-Mar 1996)
• National council for science and technology fellowship (NCST), Ministry of Education, Bangladesh (July 1991-June 1992)
• College and University merit scholarship, Ministry of Education, Bangladesh (1980-1985)

EDUCATION:

• Ph.D. 1996 Biochemistry, Division of Biological & Environmental Science Confidential, Japan 
• MS 1985 Biochemistry Confidential, Bangladesh 
• BS 1984 Biochemistry Confidential, Bangladesh