# Core B: Molecular Biology, Protein Expression and Structural Biology

> **NIH NIH P01** · CHILDREN'S HOSP OF PHILADELPHIA · 2021 · $655,208

## Abstract

Core B Abstract
The molecular biology, protein expression and structural biology core is organized around the pooling of the
resources and expertise of the project leaders in the program. The conduct of this core is based on a model
that has been successfully implemented in the past 10 years and has been extremely productive. This
centralized resource provides service and assistance in molecular biology, bacterial/yeast expression and in
the preparation of stable mammalian cell lines over-expressing vitamin K-dependent proteins, factor V variants
and factor VIII variants. The core provides the facilities, hardware and assistance for large-scale protein
production runs with stable or transiently transfected mammalian cells in serum-free conditioned media. The
core will provide facilities for large scale isolation of recombinant proteins, the appropriate chromatography
strategies and will maintain the affinity columns necessary for this activity. The core will also provide some
quality control services particularly the quantitation of -carboxyglutamic acid content to verify the integrity of
expressed vitamin K-dependent proteins. Assistance and facilities will also be provided for the large-scale
isolation of coagulation proteins from plasma. Core B personnel will assist program members in implementing
sparse matrix crystal screens, liaise with the UPenn x-ray facility to allow the collection of diffraction data on
the home source and provide expertise for optimization of crystallization conditions with expanded screens.
The core supports high performance computers equipped with necessary programs for processing diffraction
data, model building by molecular replacement and refinement. The core will also provide support for small
angle x-ray scattering measurements on the Home source and data analysis. Hydrogen/deuterium exchange
studies will be performed in collaboration with Renhao Li and facilitated by a modest sub-contract to Emory
University. The Director will be responsible for the oversight and efficient conduct of these activities and for
ensuring that equitable service is provided to the projects

## Key facts

- **NIH application ID:** 10175002
- **Project number:** 5P01HL139420-04
- **Recipient organization:** CHILDREN'S HOSP OF PHILADELPHIA
- **Principal Investigator:** Sriram Krishnaswamy
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $655,208
- **Award type:** 5
- **Project period:** 2018-09-01 → 2023-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10175002

## Citation

> US National Institutes of Health, RePORTER application 10175002, Core B: Molecular Biology, Protein Expression and Structural Biology (5P01HL139420-04). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10175002. Licensed CC0.

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