Isolation and characterization of immunoglobulin A (IgA) from bovine nasal secretions
Affiliation:
1. Heinrich-Heine-Universität Düsseldorf, Department of Rheumatology, Moorenstr. 5, 40225 Düsseldorf, Germany;2. Westfälische Wilhelms-Universität Münster, PharmaCampus, Institute of Pharmaceutical and Medicinal Chemistry, Corrensstr. 48, 48149 Münster, Germany;3. Westfälische Wilhelms-Universität Münster, Core Unit Proteomics, Interdisciplinary Center for Clinical Research, Röntgenstr. 21, 48149 Münster, Germany;4. Heinrich-Heine-Universität Düsseldorf, Department of Gynecology, Moorenstr. 5, 40225 Düsseldorf, Germany;1. Biotechnology Division, Defence Research & Development Establishment, Gwalior, 474002, India;2. Sciex, 121 DHR, Udyog Vihar, Gurugram, Haryana, India;3. School of Studies in Biotechnology, Jiwaji University, Gwalior, India;1. Laboratorio de Bioquímica, Departamento de Biociencias, Facultad de Química, UdelaR, Gral. Flores 2124, Montevideo, Uruguay;2. Laboratorio de Inmunomodulación y desarrollo de Vacunas, Departamento de Inmunobiología, Facultad de Medicina, UdelaR, Gral Flores 2125, Montevideo, Uruguay;3. Department of Molecular Cell Biology and Immunology, VU University Medical Center, De Boelelaan 1108, 1081 HZ, Amsterdam, Netherlands;1. Institute for biological research “Siniša Stanković”, University of Belgrade, Despota Stefana 142, Belgrade, Serbia;2. Institute for Multidisciplinary Research, University of Belgrade, Belgrade, Serbia;3. OMI Institute, Trnovska 8, Ljubljana, Slovenia;4. Institute of Pathophysiology, University of Ljubljana, Vrazov trg 2, Ljubljana, Slovenia;5. Institute of Mental Health, School of Medicine, University of Belgrade, Belgrade, Serbia
Abstract:
IgA was found to exist in two polymeric forms (15·3S and 12·2S) in bovine nasal secretions. The 15·3S secretory IgA (S-IgA)2 dissociated into 12·2S and 7S components and a trailing edge upon treatment with 4M Guanidine-HCl. The partial specific volume (v role=presentation style=font-size: 90%; display: inline-block; position: relative;>) and molecular weight of the 12·2S IgA were calculated to be 0·721 g/cc and 430,380 g/mole respectively. The mol. wt of the 15·3S polymer was determined to be 642,000 g per mole assuming a v role=presentation style=font-size: 90%; display: inline-block; position: relative;> of 0·721 g/cc. The 12·2S S-IgA was found to show complete identity with the 15·3S S-IgA and in addition shared antigenic determinants with human colostral IgA.J-chain was released from both species of S-IgA following treatment with 2-mercaptoethanol. The molecular weight of J-chain from S-IgA (Jα) was determined to be 17,040 by SDS-PAGE. Antigenic analysis showed Jα and Jμ (bound-form) to be identical. Stoichiometric studies of J-chain determined from densitometric scans of polyacrylamide gels of reduced and alkylated bovine 12·2S S-IgA revealed the presence of one J-chain per S-IgA molecule.
