Improvement of Biological Product Quality by Application of New Technologies to Characterize of Vaccines and Blood Products: NMR Spectroscopy and Light Scattering

Principal Investigator: Daron I. Freedberg, PhD
Office / Division / Lab: OVRR / DBPAP / LB

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Overview

Public Health Issue: Complex biological products are difficult to characterize, making the manufacture of a consistent and predictable product more challenging. Incomplete characterization of vaccines and biologics increases the risk of inconsistencies in manufacture. These risks are especially high for novel products, where traditional methods may miss subtle and unanticipated variability in products. Biophysical methods such as NMR spectroscopy and light scattering provide an invaluable complement to the biological assays traditionally used to characterize these products. For example, small amounts of free polysaccharide in conjugate vaccine formulations that may greatly reduce the efficacy of these vaccines are most effectively detected with spectroscopic methods.

Regulatory Contribution: Development and evaluation of improved tests for licensed and candidate vaccine and biologics characterization using NMR spectroscopy and light scattering will improve the ability to consistently manufacture complex biological products.

Research Approach: Development and evaluation of new methods to characterize bacterial vaccines by NMR spectroscopy, e.g., the detection of small amounts of free polysaccharide in conjugate vaccine formulations and precise measurement of vaccine and adjuvant structure and evaluation of contaminants in vaccine products.

Mission Relevance & Outcomes: Testing of vaccines and other biologics and development of new technologies for physical characterization of complex biological products will aid in evaluating licensed vaccines and candidate vaccines as well as excipients such as adjuvants. For example, recently an adulterated vaccine was identified by NMR testing.

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Publications

Curr Protoc Nucleic Acid Chem 2008 Dec;Chapter 3:Unit3.17
Release of DNA oligonucleotides and their conjugates from controlled-pore glass under thermolytic conditions.
Grajkowski A, Cie lak J, Norris S, Freedberg DI, Kauffman JS, Duff RJ, Beaucage SL

Nat Struct Mol Biol 2008 Aug;15(8):868-9
NMR structure of chaperone Chz1 complexed with histones H2A.Z-H2B.
Zhou Z, Feng H, Hansen DF, Kato H, Luk E, Freedberg DI, Kay LE, Wu C, Bai Y

Bioconjug Chem 2008 Aug;19(8):1696-706
Thermolytic release of covalently linked DNA oligonucleotides and their conjugates from controlled-pore glass at near neutral pH.
Grajkowski A, Cieslak J, Kauffman JS, Duff RJ, Norris S, Freedberg DI, Beaucage SL

Proc Natl Acad Sci U S A 2007 Jul 10;104(28):11557-61
Extracellular structure of polysialic acid explored by on cell solution NMR.
Azurmendi HF, Vionnet J, Wrightson L, Trinh LB, Shiloach J, Freedberg DI

Anal Chem 2006 Jul 1;78(13):4634-4641
Chemical Characterization of Diaspirin Cross-Linked Hemoglobin Polymerized with Poly(ethylene glycol).
Buehler PW, Boykins RA, Norris S, Alayash AI

Dev Biol 2005;122:77-83
Using nuclear magnetic resonance spectroscopy to characterize biologicals.
Freedberg DI

Anal Chem 2005 Jun 1;77(11):3466-3478
Structural and Functional Characterization of Glutaraldehyde-Polymerized Bovine Hemoglobin and Its Isolated Fractions.
Buehler PW, Boykins RA, Jia Y, Norris S, Freedberg DI, Alayash AI

Carbohydr Res 2005 Apr 11;340(5):863-74
The utility of residual dipolar couplings in detecting motion in carbohydrates: application to sucrose.
Venable RM, Delaglio F, Norris SE, Freedberg DI

Proc Natl Acad Sci U S A 2005 Apr 12;102(15):5564-9
Escherichia coli K1 polysialic acid O-acetyltransferase gene, neuO, and the mechanism of capsule form variation involving a mobile contingency locus.
Deszo EL, Steenbergen SM, Freedberg DI, Vimr ER

J Am Chem Soc 2004 Aug 25;126(33):10478-84
Discriminating the helical forms of peptides by NMR and molecular dynamics simulation.
Freedberg DI, Venable RM, Rossi A, Bull TE, Pastor RW