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NIH Research Festival

September 12 – 14, 2018

A Tandem Chromatography Approach to Determine Titer for In-Process Production of Recombinant Tetanus Toxoid Fragment C

Wednesday, September 12, 2018 – Poster Session I
12:00 – 1:30 p.m.

FAES Terrace

VRC

RSCHSUPP-3

Authors

  • N Barefoot
  • Y Yang
  • A Vinitsky
  • T Cozine
  • P Lei

Abstract

Conjugated vaccines are widely used today for treatment of a variety of diseases and represent a significant step forward in vaccine rational design. Currently the Vaccine Research Center is developing one such product for HIV-1 based on the HIV-1 envelope glycoprotein fusion peptide sequence conjugated to a recombinant tetanus toxoid fragment C (rTTHc) core. The rTTHc is produced using an E. coli based production scheme and purified prior to conjugation. Here we present a novel liquid chromatography-based approach for reporting rTTHc titer of in-process samples suitable for use immediately after cell lysis. This method relies on the sequential coupling of two different chromatography techniques in order to achieve separation; cation exchange chromatography (CEX) and size exclusion chromatography (SEC). The analyte of interest in this CEX-SEC approach, rTTHc, is separated from an extremely complex matrix consisting of host cell proteins (HCP), host cell DNA (HCD), and product related impurities such as fragment proteins in a method that allows for reliable and accurate titer determination. This novel tandem chromatography approach allows for analysis of samples from upstream and downstream development providing both teams with invaluable data for optimization of their processes. The method has been shown to be precise, specific, and repeatable.

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This page was last updated on Friday, March 26, 2021

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Current Research Festival

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  • 2024
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    • Special Wednesday Events
    • Vendor Exhibit Information
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    • Research Festival Committee
  • 2023
    • General Schedule of Events
    • NIH Early–Career Investigator Lectures
    • Poster Sessions
    • Concurrent Workshops
    • NIH Resource Information Fair
    • Vendor Exhibit Information
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  • 2021
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    • General Schedule of Events
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    • Poster Session
    • Data Blitz
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    • Special Exhibits on Resources for Intramural Research
    • Technical Sales Association (TSA) Research Festival Exhibit Tent Show
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  • 2018
    • General Schedule of Events
    • Plenary Sessions
    • Concurrent Symposia Sessions
    • Poster Sessions
    • FARE Award Ceremony
    • Special Exhibits on Resources for Intramural Research
    • Technical Sales Association (TSA) Research Festival Exhibit Tent Show
    • Research Festival Committees
  • 2017
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    • Plenary Sessions
    • Concurrent Symposia Sessions
    • Poster Sessions
    • FARE Award Ceremony
    • Future Research Leaders
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    • Technical Sales Association (TSA) Research Festival Exhibit Tent Show
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  • 2016
    • General Schedule of Events
    • Plenary Sessions
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    • Browse by Presenter Name
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    • Research Festival Committees
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