Detection of Glycoprotein Receptors on Blotting Membranes by Binding of Live Bacteria and Amplification by Growth

✍ Scribed by A. Karlsson; M. Markfjall; H. Lundqvist; N. Stromberg; C. Dahlgren

• Publisher: Elsevier Science
• Year: 1995
• Tongue: English
• Weight: 622 KB
• Volume: 224
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.1995.1055

✦ Synopsis

Conditions have been adapted for detecting bacteria bound to glycoprotein receptors on blotting membranes using a self-enhancing detection method based on bacterial growth. Neutrophil plasma membrane proteins, mediating adherence of mannose-binding type-1-fimbriated Escherichia coli and concanavalin A (Con A) to intact human neutrophils, were separated by SDSPAGE and transferred onto a polyvinylidene difluoride (PVDF) membrane. The PVDF membrane was immersed in a suspension of mannose-binding type-1fimbriated (E). coli, and after repeated washings, bound bacteria were allowed to multiply into bacterial colonies by placing the membrane on a solid nutrient substratum. About one major and eight minor glycoproteins, some of which also were detected by Con A, selectively induced colony formation in a mannose-inhibitable fashion. Binding of (\left[{ }^{35} S\right]) methionine metabolically labeled (E). coli to PVDF membranes produced a virtually identical binding pattern, demonstrating further the accuracy of this self-enhancing detection method which is rapid, simple, and sensitive and avoids radioisotopes. 1995 Academic Press, Inc.