Quantification of Antibody Persistence for Cell Surface Protein Labeling.

Cellular and Molecular Bioengineering
Megan E DempseyEric M Darling

Abstract

Antibodies are an essential research tool for labeling surface proteins but can potentially influence the behavior of proteins and cells to which they bind. Because of this, researchers and clinicians are interested in the persistence of these antibodies, particularly for live-cell applications. We developed an easily adoptable method for researchers to characterize antibody removal timelines for any cell-antibody combination, with the benefit of studying broad, hypothesized mechanisms of antibody removal. We developed a method using four experimental conditions to elucidate the contributions of possible factors influencing antibody removal: cell proliferation, internalization, permanent dissociation, and environmental perturbation. This method was tested on adipose-derived stem cells and a human lung fibroblast cell line with anti-CD44, CD90, and CD105 antibodies. The persistence of the primary antibody was probed using a fluorescent secondary antibody daily over 10 days. Relative contributions by the antibody removal mechanisms were quantified based on differences between the four culture conditions. Greater than 90% of each antibody tested was no longer present on the surface of the two cell types after 5 days, with removal ...Continue Reading

References

Feb 1, 1991·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·J T Mason, T J O'Leary
Dec 1, 1990·Pharmaceutical Research·W JiskootD J Crommelin
Dec 1, 1987·Archives of Surgery·H F SearsH Koprowski
Feb 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·I S Trowbridge, F Lopez
Apr 1, 1980·The Biochemical Journal·D W Mason, A F Williams
Jan 1, 1994·Journal of Immunotherapy with Emphasis on Tumor Immunology : Official Journal of the Society for Biological Therapy·M B KhazaeliA F LoBuglio
May 1, 1994·International Journal of Cancer. Journal International Du Cancer·I Novak-HoferP A Schubiger
Sep 27, 1993·Journal of Immunological Methods·R AlonM Wilchek
Dec 15, 1995·Journal of Immunological Methods·R AudranL Toujas
Sep 1, 1996·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·D E MosedaleD J Grainger
Sep 27, 2002·Biophysical Journal·Simone KulinKristian Helmerson
Oct 4, 2006·BMC Immunology·Ulrik B NielsenJames D Marks
Feb 2, 2008·Journal of Pharmaceutical Sciences·Sylvia KieseHanns-Christian Mahler
Jul 23, 2008·Molecular Cancer Therapeutics·Margaret E AckermanK Dane Wittrup
Apr 14, 2009·Cytometry. Part B, Clinical Cytometry·Barbara RodaGian Paolo Bagnara
Apr 27, 2010·Analytical and Bioanalytical Chemistry·Daniel R GossettDino Di Carlo
Aug 9, 2011·Experimental and Molecular Pathology·Christine A St PierreRobert W Finberg
Mar 20, 2014·Biotechnology Journal·Angela R JonesEric V Shusta
May 3, 2016·Current Stem Cell Research & Therapy·Alexander Mildmay-White, Wasim Khan
Nov 20, 2016·Annual Review of Physiology·Elizabeth A SpechtAmy E Palmer
Feb 17, 2018·Nature Communications·Toby MathiesonMikhail M Savitski
Aug 30, 2019·Journal of Pharmaceutical Sciences·Yoann Le BasleValérie Sautou
Jul 21, 2020·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Sylvia A SarnikEric M Darling

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