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 journalArticlepeer-review

63 Scopus citations

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

ASJC Scopus subject areas

  • Cell Biology

Fingerprint Dive into the research topics of 'An ELISA method for the quantitation of tracheal mucins from human and nonhuman primates.'. Together they form a unique fingerprint.

Cite this