Surface analysis of pure and complex mucin coatings
2009 (English)In: Journal of Colloid and Interface Science, ISSN 0021-9797, E-ISSN 1095-7103, Vol. 333, no 1, 180-187 p.Article in journal (Refereed) Published
In the past, we introduced the idea of using mucin coatings to improve biomaterials performance. Here, we evaluate non-radioactive methods for the analysis of pure and human host protein-containing (complex) mucin coatings on a real-type substrate (Thermanox). A common protein quantification assay (mBCA) was combined with mass-calibrated, enzyme-amplified assays based on lectin (ELLA) and antibody (ELISA) recognition, to determine the total and specific amounts of surface-associated proteins. Model studies showed the mBCA assay to be of limited use at low mass loads, and steric effects to influence the ELLA at high surface layer densities. Non-specific responses due to substrate interaction were low for the ELLA and ELISAs. Cross-reactions were observed during ELLA analysis of analytes sharing high degree of O-glycosylation. Combined mBCA-ELLA-ELISA analysis suggested that mucin desorption was low upon protein addition and that low concentrations of ELISA-determined Protein for the complex coatings Could be explained in terms of low accessibility of proteins to the bulk environment. Specifically, a methodology is presented for the determination of the fraction of surface-exposed, presumed bioactive proteins in a complex mucin coating. Finally, X-ray photoelectron spectroscopy and infrared reflectance spectroscopy combined with multivariate data analysis were proven useful in the evaluation of mucin-based coatings.
Place, publisher, year, edition, pages
2009. Vol. 333, no 1, 180-187 p.
Mucin quantification, mBCA, ELLA, ELISA, Ellipsometry, Jacalin, Complex mucin coating, Surface-exposed protein, Bioactivity, XPS, IRS-MVDA, Principal component analysis (PCA)
IdentifiersURN: urn:nbn:se:uu:diva-97900DOI: 10.1016/j.jcis.2009.01.069ISI: 000264897200024OAI: oai:DiVA.org:uu-97900DiVA: diva2:173005