Fnu Shahid, Ph.D. PMP

Fnu Shahid, Ph.D. PMP

Princeton, New Jersey, United States
2K followers 500+ connections

About

Specializing in Cellular and Gene Therapies (CGT) using CAR (Chimeric Antigen Receptor)…

Contributions

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Experience

  • BioCentriq Graphic

    BioCentriq

    New Jersey, United States

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    Webster, Texas, United States

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    Bethesda, Maryland, United States

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    Narayanganj, Dhaka, Bangladesh

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    Dhaka Bangladesh

Education

  • University of Wisconsin-Milwaukee Graphic

    University of Wisconsin-Milwaukee

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    Selected Course Studied:
    *Protein Structure and Function
    *Biochemistry-Cellular Processes and Metabolism
    *Molecular Biology-Microorganism
    *Chemical Kinetics and Dynamics
    *Liquid Chromatography-Mass Spectrometry Fundamentals and Applications
    *Medicinal Chemistry: Drug Discovery/Lead Optimization & DNA as Drug Targets

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    Selected Course Studied:
    *Advanced Molecular Biology
    *Bioinformatics
    *Biochemistry of Drugs
    *Molecular Diagnostics and DNA Forensic Sciences
    *Environmental Biochemistry
    *Applied Nutrition and Nutritional Biochemistry

    Master's Thesis Topic: Effects of Vitamin B12 in hemolytic anemia in mother and children.
    During my research
    1) I was trained to do serological and clinical immunology tests, ELISA, RT-PCR, ALS and etc for multiple projects funded by UC-DAVIS.
    2)…

    Selected Course Studied:
    *Advanced Molecular Biology
    *Bioinformatics
    *Biochemistry of Drugs
    *Molecular Diagnostics and DNA Forensic Sciences
    *Environmental Biochemistry
    *Applied Nutrition and Nutritional Biochemistry

    Master's Thesis Topic: Effects of Vitamin B12 in hemolytic anemia in mother and children.
    During my research
    1) I was trained to do serological and clinical immunology tests, ELISA, RT-PCR, ALS and etc for multiple projects funded by UC-DAVIS.
    2) Collected blood (and cord blood during child birth) samples from pregnant women, 4 months infants and analyzed antibodies (IgG, IgM, IgA, folic acid, Vitamin B12, etc) for placebo group and vitamin B12 supplemented group to study hemolytic anemia.

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    Undergrad Research:
    Bioinformatics analysis and homology comparison of Cox2 enzyme in various species.

    Selected Course Studied:
    * Immunology
    * Forensic DNA Analysis
    * Clinical Biochemistry
    * Microbiology
    * Biostatistics
    * Physiology
    * Carbohydrate, Lipid, and Protein Metabolism
    * Genetics
    * Organic Chemistry and Secondary Metabolites

Licenses & Certifications

Publications

  • Rapid manufacture of low-seed CAR-T cells in a GMP-grade hollow-fiber bioreactor platform

    International Society for Cell & Gene Therapy

    Expanding CD19 CAR-T cells from varying starting material amounts (1, 3, 6 and 15 million cells) with the Quantum FlexTM Cell Expansion System from Terumo Blood and Cell Technologies. Based on the use of commercially available T cells and an anti-CD19 CAR-T lentiviral construct, the system achieved a 150- to 200-fold increase in cell numbers over 7 days, yielding 2.6 billion cells from 15 million starting cells. Across all runs, cell viability remained above 93%. In order to cryopreserve the…

    Expanding CD19 CAR-T cells from varying starting material amounts (1, 3, 6 and 15 million cells) with the Quantum FlexTM Cell Expansion System from Terumo Blood and Cell Technologies. Based on the use of commercially available T cells and an anti-CD19 CAR-T lentiviral construct, the system achieved a 150- to 200-fold increase in cell numbers over 7 days, yielding 2.6 billion cells from 15 million starting cells. Across all runs, cell viability remained above 93%. In order to cryopreserve the cells, we used the FiniaTM Fill and Finish System. In a post-manufacturing analysis, the expanded CAR-T cells proved potent and cytotoxic, making them suitable for both adult and pediatric therapies. According to the study, selecting the right cell expansion platform is key to maximizing the efficiency and scalability of CAR-T production.

    See publication
  • Asthmatics with High Serum Amyloid A have Pro-inflammatory HDL: Implications for Augmented Systemic and Airway Inflammation

    Journal of Allergy and Clinical Immunology

    Aʙꜱᴛʀᴀᴄᴛ
    Serum amyloid A (SAA) is bound to high-density lipoproteins (HDL) in blood. Although SAA is increased in the blood of asthmatics, it is not known whether this modifies asthma severity.

    Objective
    To define the clinical characteristics of asthmatics with high SAA levels and assess whether HDL from SAA-high asthmatics is pro-inflammatory.

    Methods
    SAA levels in serum from asthmatic and non-asthmatic subjects were quantified by ELISA. HDL isolated from asthmatics with…

    Aʙꜱᴛʀᴀᴄᴛ
    Serum amyloid A (SAA) is bound to high-density lipoproteins (HDL) in blood. Although SAA is increased in the blood of asthmatics, it is not known whether this modifies asthma severity.

    Objective
    To define the clinical characteristics of asthmatics with high SAA levels and assess whether HDL from SAA-high asthmatics is pro-inflammatory.

    Methods
    SAA levels in serum from asthmatic and non-asthmatic subjects were quantified by ELISA. HDL isolated from asthmatics with high SAA levels were used to stimulate human monocytes and were intravenously administered to BALB/c mice.

    Results
    A SAA level > 108.8 μg/ml was defined as the threshold to identify 11% of an asthmatic cohort (n = 146) as being SAA-high. SAA-high asthmatics were characterized by increased serum C-reactive protein, IL-6, and TNF-α; older age; and an increased prevalence of obesity and severe asthma. HDL isolated from SAA-high asthmatics (SAA-high HDL) had an increased content of SAA as compared to HDL from SAA-low asthmatics and induced the secretion of IL-6, IL-1β and TNF-α from human monocytes via a FPR2/ATP/P2X7R axis. Intravenous administration to mice of SAA-high HDL, but not normal HDL, induced systemic inflammation and amplified allergen-induced neutrophilic airway inflammation and goblet cell metaplasia.

    Conclusion
    SAA-high asthmatics are characterized by systemic inflammation, older age, and an increased prevalence of obesity and severe asthma. HDL from SAA-high asthmatics is pro-inflammatory and, when intravenously administered to mice, induces systemic inflammation, and amplifies allergen-induced neutrophilic airway inflammation. This suggests that systemic inflammation induced by SAA-high HDL may augment disease severity in asthma.

    See publication
  • A Kinetic Investigation of the Early Steps in Cytochrome c Nitrite Reductase (ccNiR)-Catalyzed Reduction of Nitrite

    Biochemistry

    The decaheme enzyme cytochrome c nitrite reductase (ccNiR) catalyzes reduction of nitrite to ammonium in a six-electron, eight-proton process. With a strong reductant as the electron source, ammonium is the sole product. However, intermediates accumulate when weaker reductants are employed, facilitating study of the ccNiR mechanism. Herein, the early stages of Shewanella oneidensis ccNiR-catalyzed nitrite reduction were investigated by using the weak reductants…

    The decaheme enzyme cytochrome c nitrite reductase (ccNiR) catalyzes reduction of nitrite to ammonium in a six-electron, eight-proton process. With a strong reductant as the electron source, ammonium is the sole product. However, intermediates accumulate when weaker reductants are employed, facilitating study of the ccNiR mechanism. Herein, the early stages of Shewanella oneidensis ccNiR-catalyzed nitrite reduction were investigated by using the weak reductants N,N,N′,N’-tetramethyl-p-phenylenediamine (TMPD) and ferrocyanide. In stopped-flow experiments, reduction of nitrite-loaded ccNiR by TMPD generated a transient intermediate, identified as FeH1II(NO2–), where FeH1 represents the ccNiR active site. FeH1II(NO2–) accumulated rapidly and was then more slowly converted to the two-electron-reduced moiety {FeH1NO}7; ccNiR was not reduced beyond the {FeH1NO}7 state. The midpoint potentials for sequential reduction of FeH1III(NO2–) to FeH1II(NO2–) and then to {FeH1NO}7 were estimated to be 130 and 370 mV versus the standard hydrogen electrode, respectively. FeH1II(NO2–) does not accumulate at equilibrium because its reduction to {FeH1NO}7 is so much easier than the reduction of FeH1III(NO2–) to FeH1II(NO2–). With weak reductants, free NO• was released from nitrite-loaded ccNiR. The release of NO• from {FeH1NO}7 is exceedingly slow (k ∼ 0.001 s–1), but it is somewhat faster (k ∼ 0.050 s–1) while FeH1III(NO2–) is being reduced to {FeH1NO}7; then, the release of NO• from the undetectable transient {FeH1NO}6 can compete with reduction of {FeH1NO}6 to {FeH1NO}7. CcNiR appears to be optimized to capture nitrite and minimize the release of free NO•. Nitrite capture is achieved by reducing bound nitrite with even weak electron donors, while NO• release is minimized by stabilizing the substitutionally inert {FeH1NO}7 over the more labile {FeH1NO}6.

    See publication
  • New Mechanistic Insights about the Enzyme Cytochrome c nitrite reductase (ccNiR) from studies of the wild type and its variants

    The FASEB Journal

    Cytochrome c Nitrite Reductase (ccNiR) is a periplasmic, decaheme homodimeric enzyme that catalyzes the six-electron reduction of nitrite to ammonia. Under standard assay conditions catalysis proceeds without detected intermediates, and it has been assumed that this is also true in vivo. However, in vitro we have found it possible to trap putative intermediates, or to release partially reduced nitrogen species such as nitric oxide, by controlling the electrochemical potential at which reduction…

    Cytochrome c Nitrite Reductase (ccNiR) is a periplasmic, decaheme homodimeric enzyme that catalyzes the six-electron reduction of nitrite to ammonia. Under standard assay conditions catalysis proceeds without detected intermediates, and it has been assumed that this is also true in vivo. However, in vitro we have found it possible to trap putative intermediates, or to release partially reduced nitrogen species such as nitric oxide, by controlling the electrochemical potential at which reduction takes place. This poster will present UV-Vis spectropotentiometric titrations of the active site variants R103Q and H257Q, and steady-state and stopped-flow kinetic studies of ccNiR-catalyzed reduction of nitrite to nitric oxide by the weak reductant N,N,N′,N′-tetramethyl-p-phenylenediamine (TMPD). Wild type nitrite-loaded ccNiR is reduced chemically or electrochemically at very high potentials (midpoint potential of 246 mV vs SHE) in a concerted 2-electron process that forms an {FeNO}7 moiety at the active site. By contrast, the H257Q variant’s active site midpoint potential is downshifted to 125 mV vs SHE and is now a 1-electron reduction to yield an {FeNO}6 moiety. The wild type catalyzes the reduction of nitrite to NO• by TMPD. H257Q does not appear to catalyze this reaction, which may be due to its lower active site midpoint potential or to the much slower turnover of the mutant; studies with more powerful reductants are now under way

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  • Trapping of a Putative Intermediate in the Cytochrome c Nitrite Reductase (ccNiR)-Catalyzed Reduction of Nitrite: Implications for the ccNiR Reaction Mechanism

    Journal of the American Chemical Society (JACS)

    Cytochrome c nitrite reductase (ccNiR) is a periplasmic, decaheme homodimeric enzyme that catalyzes the six-electron reduction of nitrite to ammonia. Under standard assay conditions catalysis proceeds without detected intermediates, and it has been assumed that this is also true in vivo. However, this report demonstrates that it is possible to trap a putative intermediate by controlling the electrochemical potential at which reduction takes place. UV/vis spectropotentiometry showed that…

    Cytochrome c nitrite reductase (ccNiR) is a periplasmic, decaheme homodimeric enzyme that catalyzes the six-electron reduction of nitrite to ammonia. Under standard assay conditions catalysis proceeds without detected intermediates, and it has been assumed that this is also true in vivo. However, this report demonstrates that it is possible to trap a putative intermediate by controlling the electrochemical potential at which reduction takes place. UV/vis spectropotentiometry showed that nitrite-loaded Shewanella oneidensis ccNiR is reduced in a concerted two-electron step to generate an {FeNO}7 moiety at the active site, with an associated midpoint potential of +246 mV vs SHE at pH 7. By contrast, cyanide-bound active site reduction is a one-electron process with a midpoint potential of +20 mV, and without a strong-field ligand the active site midpoint potential shifts 70 mV lower still. EPR analysis subsequently revealed that the {FeNO}7 moiety possesses an unusual spectral signature, different from those normally observed for {FeNO}7 hemes, that may indicate magnetic interaction of the active site with nearby hemes. Protein film voltammetry experiments previously showed that catalytic nitrite reduction to ammonia by S. oneidensis ccNiR requires an applied potential of at least −120 mV, well below the midpoint potential for {FeNO}7 formation. Thus, it appears that an {FeNO}7 active site is a catalytic intermediate in the ccNiR-mediated reduction of nitrite to ammonia, whose degree of accumulation depends exclusively on the applied potential. At low potentials the species is rapidly reduced and does not accumulate, while at higher potentials it is trapped, thus preventing catalytic ammonia formation.

    See publication

Honors & Awards

  • Chancellor’s graduate fellowship from the Department of Chemistry and Biochemistry

    Department of Chemistry and Biochemistry

Organizations

  • Bangladesh Student Association

    President

    - Present

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