High-fidelity and iterative affinity extraction of hyaluronan.

Autor: Erxleben DA; Virginia Tech-Wake Forest University School of Biomedical Engineering and Sciences, Wake Forest University School of Medicine Winston-Salem North Carolina USA., Rivas F; Virginia Tech-Wake Forest University School of Biomedical Engineering and Sciences, Wake Forest University School of Medicine Winston-Salem North Carolina USA., Smith I; Department of Biology Wake Forest University Winston-Salem North Carolina USA., Poddar S; Virginia Tech-Wake Forest University School of Biomedical Engineering and Sciences, Wake Forest University School of Medicine Winston-Salem North Carolina USA., DeAngelis PL; Department of Biochemistry and Physiology University of Oklahoma Health Sciences Center Oklahoma City Oklahoma USA., Rahbar E; Departments of Biomedical Engineering and Veterinary Physiology and Pharmacology Texas A&M University College Station Texas USA., Hall AR; Virginia Tech-Wake Forest University School of Biomedical Engineering and Sciences, Wake Forest University School of Medicine Winston-Salem North Carolina USA.; Comprehensive Cancer Center, Wake Forest University School of Medicine Winston-Salem North Carolina USA.
Jazyk: angličtina
Zdroj: Proteoglycan research [Proteoglycan Res] 2024 Oct-Dec; Vol. 2 (4), pp. e70008. Date of Electronic Publication: 2024 Dec 06.
DOI: 10.1002/pgr2.70008
Abstrakt: The glycosaminoglycan hyaluronan (HA) serves a variety of crucial physiological functions in vertebrates. Synthesized at the plasma membrane and secreted into the extracellular environment, HA polymers span a wide range of molecular weights (MW) that define their activity through a notable size-function relationship. Analytical technologies for determining HA MW distributions typically require selective extraction from complex biofluids or tissues. A common method for achieving this is immunoprecipitation-like pull-down using specific HA-binding proteins bound to magnetic beads. Here, we present a systematic investigation of experimental variables involved in this process, leading to an affinity extraction protocol that enables iterative bead reuse and reagent lifetime maximization, thereby enhancing the efficiency of the HA extraction process. Our methods provide a framework for general optimization of immunoprecipitation in other contexts with heterogenous analyte sizes.
Competing Interests: A.R.H, P.L.D, and E.R are listed as inventors on a patent covering SSNP analysis of HA.
(© 2024 The Author(s). Proteoglycan Research published by Wiley Periodicals LLC.)
Databáze: MEDLINE
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