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PMID: 6778926 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Use of Pseudomonas aeruginosa lipopolysaccharide immunoadsorbents to prepare high potency, mono-specific antibodies.

Journal of immunological methods ·Vol. 38 ·No. 1-2 ·1980-00-00 ·Pages 103-16

Fick RB, Naegel GP, Reynolds HY

Abstract

Potent, mono-specific anti-Pseudomonas immunoglobulins were isolated from serum and lung lavage fluid from patients with cystic fibrosis using immunotype specific Pseudomonas aeruginosa lipopolysaccharide (LPS) substituted immunoadsorbent gel. Iodinated monovalent Pseudomonas LPS somatic antigens, Fisher immunotypes, were used as ligands for two different insoluble gel matrices. LPS iodination, using the water insoluble chloroglycoluril reagent, permitted quantitation of the percent LPS bound as a ligand. The coupling efficiencies of epoxy-activated and cyanogen bromide-activated Sepharose matrices for various Pseudomonas immunotype specific LPS preparations were compared. Although each of the 7 LPS somatic antigens produced an equivalent amount of coupling, higher percentages of coupling were found using the cyanogen bromide-activated gel when compared to the epoxy-activated gel. IgG fractions prepared from cystic fibrosis sera and lung lavage fluid were passed through the LPS affinity gels, and Pseudomonas LPS immunotype specific antibodies were eluted. The presence of specific antibody activity against individual Pseudomonas immunotypes was determined with a passive micro-hemagglutination assay. Bronchial lavage fluid seemed to be as effective as serum as a source of Pseudomonas specific antibody. Use of such a LPS substituted gel permits direct recovery of Pseudomonas monospecific antibodies suitable for physical-chemical analyses and for biologic functional assays.

MeSH Terms
Antibodies, Bacterial/isolation & purification Body Fluids/immunology Cystic Fibrosis/immunology Humans Immunoglobulin G/immunology Immunosorbent Techniques Lipopolysaccharides/immunology Lung/immunology Pseudomonas aeruginosa/immunology
Chemicals
Antibodies, Bacterial Immunoglobulin G Lipopolysaccharides
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Fick R B
Naegel G P
Reynolds H Y
Article Info
Journal
Journal of immunological methods
Abbr.
J Immunol Methods
ISSN
0022-1759
Published
1980-00-00
Pages
103-16
Language
English
Region
Netherlands
NLM ID
1305440
Subset
IM
Grants
PHS HHS · 22302L · United States
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