An ELISA method for the quantitation of tracheal mucins from human and nonhuman primates.

H. Lin, D. M. Carlson, J. A. St George, Charles Plopper, Reen Wu

Research output: Contribution to journalArticle

62 Citations (Scopus)

Abstract

Monoclonal antibodies, 17B1 and 17Q2, which are specific for large molecular weight mucous glycoproteins of airway epithelium, have been used to develop an ELISA method to quantitate the tracheal mucins of humans and rhesus monkeys. The assay is a double-sandwich system that does not depend on either the binding of mucous antigens to the microtiter plate or the use of a second antibody. The assay protocol includes (1) coating the microtiter well with purified IgG of 17B1 or 17Q2, (2) incubating the wells with mucous samples, (3) binding of alkaline phosphatase-conjugated IgG to the wells, and (4) developing the color with phosphate substrate. This ELISA method is very sensitive for human and rhesus monkey tracheal mucins. Quantitation is not affected by the presence of various proteoglycans (keratan sulfate, hyaluronate, heparin, heparan sulfate, and chondroitin sulfate). However, the quantitation is affected by the treatment of antigen with periodic acid and endo-beta-galactosidase. Other enzymes (e.g., neuraminidase, hyaluronidase, chondroitinase, heparitinase, heparinase, fucosidase, keratanase) have no effect on the antigenicity of substrate. The quantitation is linear, with a concentration from 0.2 to 4 ng protein/sample. The ELISA method developed in this study should be useful for quantitating the mucin content of various biologic fluids, such as sputum, bronchoalveolar lavage, and media from cultures following various pharmacologic and physiologic manipulations.

Original languageEnglish (US)
Pages (from-to)41-48
Number of pages8
JournalAmerican Journal of Respiratory Cell and Molecular Biology
Volume1
Issue number1
StatePublished - Jul 1989

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keratan-sulfate endo-1,4-beta-galactosidase
Mucins
Primates
Enzyme-Linked Immunosorbent Assay
heparitinsulfate lyase
Macaca mulatta
Assays
Immunoglobulin G
Chondroitinases and Chondroitin Lyases
Heparin Lyase
alpha-L-Fucosidase
Keratan Sulfate
Antigens
Periodic Acid
Hyaluronoglucosaminidase
Heparitin Sulfate
Chondroitin Sulfates
Neuraminidase
Bronchoalveolar Lavage
Proteoglycans

ASJC Scopus subject areas

  • Cell Biology

Cite this

An ELISA method for the quantitation of tracheal mucins from human and nonhuman primates. / Lin, H.; Carlson, D. M.; St George, J. A.; Plopper, Charles; Wu, Reen.

In: American Journal of Respiratory Cell and Molecular Biology, Vol. 1, No. 1, 07.1989, p. 41-48.

Research output: Contribution to journalArticle

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