Creation of the first monoclonal antibody recognizing an extracellular epitope of hABCC6. Issue 6 (28th November 2020)

• Record Type: Journal Article
• Authors: Kozák, Eszter; Szikora, Bence; Iliás, Attila; Jani, Péter K.; Hegyi, Zoltán; Matula, Zsolt; Dedinszki, Dóra; Tőkési, Natália; Fülöp, Krisztina; Pomozi, Viola; Várady, György; Bakos, Éva; Tusnády, Gabor E.; Kacskovics, Imre; Váradi, Andras
• Is Part Of: FEBS letters. Volume 595:Issue 6(2021)
• Page Start: 789
• Page End: 798
• Publication Date: 2020-11-28
• Subjects: ABCC6 -- ectopic calcification -- extracellular epitope -- monoclonal antibody; Biochemistry -- Periodicals; Biophysics -- Periodicals; Molecular biology -- Periodicals; Biochimie -- Périodiques; Biochemistry; Biophysics; Molecular biology; Periodicals; 572.05
• DOI: 10.1002/1873-3468.13991
• Languages: English
• ISSNs: 0014-5793

Abstract

Abstract : Mutations in the ABCC6 gene result in calcification diseases such as pseudoxanthoma elasticum or Generalized Arterial Calcification of Infancy. Generation of antibodies recognizing an extracellular (EC) epitope of ABCC6 has been hampered by the short EC segments of the protein. To overcome this limitation, we immunized bovine FcRn transgenic mice exhibiting an augmented humoral immune response with Human Embryonic Kidney 293 cells cells expressing human ABCC6 (hABCC6). We obtained a monoclonal antibody recognizing an EC epitope of hABCC6 that we named mEChC6. Limited proteolysis revealed that the epitope is within a loop in the N‐terminal half of ABCC6 and probably spans amino acids 338–347. mEChC6 recognizes hABCC6 in the liver of hABCC6 transgenic mice, verifying both specificity and EC binding to intact hepatocytes. Abstract :