https://pages.htgaa.org/2026b/tania-araceli-barreda-galvez/index.htmlAbout me I'm a bioengineer from UTEC (Lima, Peru) with a background that bridges wet lab research and regulatory affairs. During a research internship at the University of Alberta, I worked on cell culture, PCR, and fluorescence microscopy, and I led a fermentation kinetics project on chicha de jora funded by the Harvard-UTEC Seed Fund. I currently work at a pathology laboratory in Peru, verifying compliance of immunohistochemistry results for oncology partnerships, an experience that sparked my interest in biomanufacturing, particularly cell-free systems and automation applied to diagnostics. I'm excited to join HTBAA as a Committed Listener and explore how to translate lab-based discoveries into real, scalable products.Hugoenhttps://pages.htgaa.org/2026b/tania-araceli-barreda-galvez/week-01-hw-project-ideas/index.htmlMon, 01 Jan 0001 00:00:00 +0000https://pages.htgaa.org/2026b/tania-araceli-barreda-galvez/week-01-hw-project-ideas/index.htmlHTBAA Homework #1 1. What you would like to biomanufacture and why? Target product: a single-domain antibody fragment (nanobody, VHH) directed against PD-L1, intended for use as an immunohistochemistry (IHC) diagnostic reagent, designed to integrate with an automated digital quantification workflow. Why: In triple-negative breast cancer (TNBC), eligibility for atezolizumab depends on a specific assay, SP142, which evaluates PD-L1 expression in tumor-infiltrating immune cells using a threshold of just 1%. This type of scoring presents challenges related to the pathologist’s visual assessment and inter-pathologist agreement when evaluating low thresholds in tissue. Without prior training, visual interpretation tends to be more subjective, and while training improves outcomes, it does not eliminate reliance on individual visual interpretation (Kim et al., 2021). This substantial variability among pathologists when evaluating immunological biomarkers has direct implications for therapeutic decisions (Van Bockstal et al., 2023). I have encountered this type of problem firsthand in my current position at a pathology laboratory in Peru. The accuracy of a reagent is not merely a technical issue; rather, it determines whether a patient has access to treatment. A nanobody produced through a standardized cell-free process, combined with an automated digital quantification system rather than a pathologist’s visual reading, could offer a more objective alternative to the current manual scoring. As far as I can tell from the literature, this combination has not been specifically explored for this problem.https://pages.htgaa.org/2026b/tania-araceli-barreda-galvez/week-02-hw-dna-design/index.htmlMon, 01 Jan 0001 00:00:00 +0000https://pages.htgaa.org/2026b/tania-araceli-barreda-galvez/week-02-hw-dna-design/index.htmlHTBAA Homework #2 0. Set up in Kernel My repository is tania-barreda-PDL1 1. Define your expression target Molecule: a protein, the anti-PD-L1 nanobody (VHH) KN035. Sequence and source: the primary sequence comes from PDB entry 5JDS (chain B, “Nanobody,” Camelidae), also indexed in NCBI Structure under MMDB ID 149666, from Zhang et al. (2017). Because KN035 is an engineered antibody rather than a natural gene product, it has no dedicated UniProt entry; the target antigen, human PD-L1, does (UniProt Q9NZQ7). As a backup source for the same molecule, KN035 is also disclosed as SEQ ID NO. 1 in patent US10556954B2. The final CDS I designed adds an initiator Met before this sequence and, separately, a double stop codon (TGA-TAA) at the end, since the deposited sequence has neither.https://pages.htgaa.org/2026b/tania-araceli-barreda-galvez/week-03-hw-cell-free/index.htmlMon, 01 Jan 0001 00:00:00 +0000https://pages.htgaa.org/2026b/tania-araceli-barreda-galvez/week-03-hw-cell-free/index.htmlA. Compare cell-free reaction compositions 1. Why does the concentration of potassium glutamate and magnesium glutamate differ between compositions? Which is more sensitive? The concentrations of potassium glutamate and magnesium glutamate differ between formulations because cell-free protein synthesis (CFPS) performance depends on the overall chemical environment, so the reagent concentrations have to be optimized together rather than one at a time. The glutamate is only the counter-ion; what matters is the K⁺ and Mg²⁺ it delivers. K⁺ mainly sets the ionic strength of the reaction, while Mg²⁺ stabilizes the ribosome and is required for ATP and other nucleotide triphosphates to be usable by the enzymes. Because NTPs, ATP and the inorganic phosphate released from PEP all bind Mg²⁺, a formulation with a different energy and nucleotide system needs a different total amount of Mg²⁺ to keep the right amount free. The lysate and the fixed base buffer also contribute their own ions.https://pages.htgaa.org/2026b/tania-araceli-barreda-galvez/final-project/index.htmlMon, 01 Jan 0001 00:00:00 +0000https://pages.htgaa.org/2026b/tania-araceli-barreda-galvez/final-project/index.html