• Decrease font size
  • Return font size to normal
  • Increase font size
U.S. Department of Health and Human Services

Vaccines, Blood & Biologics

  • Print
  • Share
  • E-mail

Studies on the Safety and Efficacy of Plasma-derived Products and Their Recombinant Analogs

Principal Investigator: Basil Golding, MD
Office / Division / Lab: OBRR / DH / LPD

General Overview

The goal of our research is to enhance the safety of plasma-derived products, such as proteins used to treat clotting disorders, immune system disorders, and certain other diseases.

Our laboratory is pursuing this goal by developing tests that detect contamination of these products by substances released by microorganisms. Although microbes are filtered out of plasma used to make the products, the small molecules they make can escape filtration and trigger inflammation in individuals treated with the products.

Our tests use a variety of cells that carry molecules called toll-like receptors (TLRs) on their surface. Each group of cells carries a specific type of TLR, which detects and binds to a specific protein made by microorganisms. Tests based on this interaction between TLRs and these proteins, called TLR ligands, will help FDA and industry screen plasma-derived products for contamination by microorganisms or their products.

FDA will make any tests that we develop available to industry so they can be used to increase the safety of plasma-derived products.

Scientific Overview

Toll like receptors (TLRs) are naturally occurring pathogen-recognition receptors found on cells of the innate immune system in all species from drosophila to humans. We developed a panel of human cell lines that express TLRs that detect both human pathogens and compounds derived from these pathogens.

Plasma-derived products, such as immune globulins, albumin, and coagulation factors, are manufactured from plasma pools. Because certain parts of the manufacturing process are not performed under sterile conditions, some microbial contamination occurs. Therefore, products undergo sterile filtration of the bulk prior to being put into vials. Thus intact pathogens are removed, but compounds released from pathogens can remain in the final product. One of the commonest adverse effects of immune globulin administration is chills and fever, most likely due to presence of such compounds in the product.

Recombinant products made in bacteria or yeasts can also be contaminated with microbial compounds during the manufacturing process. Again, sterile filtration will remove the intact organisms but not the pro-inflammatory soluble compounds that can be present.

Current methods for detection of bacterial compounds are the rabbit pyrogen assay and the limulus amebocyte assay (LAL test). Both of these are based on reactivity of non-human cells and use animals.

In contrast, the method we have developed is based on the reactivity of human cells.

We established the utility of the method by showing that a recombinant product associated with serious adverse events in humans contained flagellin derived from E. coli, which was used as an expression vector. The final container testing of the product using routine methods did not detect this contaminant. In contrast, The panel of TLR cell lines that we developed showed that the product reacted with cells expressing TLR5, the ligand for flagellin. We demonstrated the specificity of this binding by inhibiting it with monoclonal anti-TLR5 antibody. We then used Western blot to show that the product contained flagellin, and confirmed that it was E. coli flagellin using mass spectrometry.


J Clin Microbiol 2009 Nov;47(11):3427-34
Use of toll-like receptor assays to detect and identify microbial contaminants in biological products. Exit Disclaimer
Huang LY, Dumontelle JL, Zolodz M, Deora A, Mozier NM, Golding B

Biotechnol Bioeng 2009 Feb 15;102(3):828-44
Production, purification, and characterization of human alpha1 proteinase inhibitor from Aspergillus niger. Exit Disclaimer
Chill L, Trinh L, Azadi P, Ishihara M, Sonon R, Karnaukhova E, Ophir Y, Golding B, Shiloach J

Biologicals 2007 Oct;35(4):285-95
Development and evaluation of an ELISA for quantification of human alpha-1-proteinase inhibitor in complex biological mixtures. Exit Disclaimer
Karnaukhova E, Golding B, Ophir Y

Microb Cell Fact 2007 Oct 29;6:34
Expression of human alpha1-proteinase inhibitor in Aspergillus niger. Exit Disclaimer
Karnaukhova E, Ophir Y, Trinh L, Dalal N, Punt PJ, Golding B, Shiloach J

Cancer Res 2007 May 15;67(10):5059
Regulatory B cells inhibit antitumor immunity. Exit Disclaimer
Inoue S, Scott D, Golding B, Leitner WW

Microbes Infect 2007 Jan;9(1):55-62
Brucella abortus bacA mutant induces greater pro-inflammatory cytokines than the wild-type parent strain. Exit Disclaimer
Parent MA, Goenka R, Murphy E, Levier K, Carreiro N, Golding B, Ferguson G, Roop RM 2nd, Walker GC, Baldwin CL

Vaccine 2006 Jul 26;24(31-32):5872-80
Passive immunotherapy of Bacillus anthracis pulmonary infection in mice with antisera produced by DNA immunization. Exit Disclaimer
Herrmann JE, Wang S, Zhang C, Panchal RG, Bavari S, Lyons CR, Lovchik JA, Golding B, Shiloach J, Lu S

Amino Acids 2006 Jun;30(4):317-32
Recombinant human alpha-1 proteinase inhibitor: towards therapeutic use. Exit Disclaimer
Karnaukhova E, Ophir Y, Golding B

Immunol Lett 2005 Sep 15;100(2):195-201
The cytotoxic T lymphocyte response against a protein antigen does not decrease the antibody response to that antigen although antigen-pulsed B cells can be targets. Exit Disclaimer
Wang W, Golding B

J Immunol 2005 Sep 15;175(6):3964-70
Th1-Like Cytokine Induction by Heat-Killed Brucella abortus Is Dependent on Triggering of TLR9. Exit Disclaimer
Huang LY, Ishii KJ, Akira S, Aliberti J, Golding B

Thromb Res 2005 2005;116(4):335-44
Identification of a novel immunodominant cytotoxic T lymphocyte epitope derived from human factor VIII in a murine model of hemophilia A. Exit Disclaimer
Wang W, Merchlinsky M, Inman J, Golding B

Vaccine 2005 Feb 25;23(14):1730-8
Programming of CTL with heat-killed Brucella abortus and antigen allows soluble antigen alone to generate effective secondary CTL. Exit Disclaimer
Inoue S, Golding B, Scott D

Clin Diagn Lab Immunol 2004 Nov;11(6):1158-64
Characterization of Antibodies to Capsular Polysaccharide Antigens of Haemophilus influenzae Type b and Streptococcus pneumoniae in Human Immune Globulin Intravenous Preparations. Exit Disclaimer
Mikolajczyk MG, Concepcion NF, Wang T, Frazier D, Golding B, Frasch CE, Scott DE

J Med Primatol 2004 Aug;33(4):167-74
Systemic and mucosal immunity in rhesus macaques immunized with HIV-1 peptide and gp120 conjugated to Brucella abortus. Exit Disclaimer
Eller N, Golding H, Inoue S, Beining P, Inman J, Matthews N, Scott DE, Golding B


Contact FDA

(800) 835-4709
(240) 402-8010
Consumer Affairs Branch (CBER)

Division of Communication and Consumer Affairs

Office of Communication, Outreach and Development

Food and Drug Administration

10903 New Hampshire Avenue

Building 71 Room 3103

Silver Spring, MD 20993-0002