Analysis of swine beta1 integrin (CD29) epitopes through monoclonal antibodies developed using two immunization strategies.
This report describes the production and characterization of monoclonal antibodies (MAbs) to swine beta1 integrin subunit (CD29) using two different immunization strategies. MAb GP4B4 was developed from a mouse immunized with porcine peripheral blood mononuclear cells (PBMC), while MAbs GP1A5, GP1A6, and GP4A1 were produced by immunizing mice with a porcine CD29 recombinant protein (rpCD29) developed in our laboratory, which includes the ligand binding site. GP4B4 and UCP1D2 (specific to porcine CD29) immunoprecipitated two bands of approximately 115 and 150 kDa under reducing conditions. The molecule recognized by these two antibodies was studied using flow cytometry and was found in practically all cells studied, displaying a similar reaction pattern. Western blot assays performed with rpCD29 indicated that MAbs GP1A5, GP1A6, and GP4A1 recognized the 30-kDa band for this recombinant protein, confirming their specificity for the beta1, integrin subunit. Immunohistochemical analyses of these MAbs disclosed a morphological pattern associated with smooth muscle, epithelium, and myeloid cells, as expected in MAbs recognizing CD29. This MAb panel could be useful as a general anti-CD29 reagent and would allow further research into this important integrin in swine.
Paños
G
G
Unidad Mixta CSIC-UCO Marcadores Genéticos Moleculares en Animales Domésticos, Departamento de Genética, Facultad de Veterinaria, Universidad de Córdoba, Córdoba, Spain. gelmoloa@uco.es
Moreno
A
A
Jiménez-Marín
A
A
Garrido
J J
JJ
Martin de la Mulas
J
J
Ordás
J
J
Llanes
D
D
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D015153:Blotting, Western; D000939:Epitopes / Q000032:analysis / Q000276:immunology*; D005434:Flow Cytometry; D006801:Humans; D007114:Immunization / Q000379:methods; D007150:Immunohistochemistry; D019012:Integrin beta1 / Q000032:analysis / Q000276:immunology*; D007963:Leukocytes, Mononuclear / Q000276:immunology*; D051379:Mice; D009928:Organ Specificity; D011233:Precipitin Tests; D021122:Protein Subunits / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology; D013045:Species Specificity; D013552:Swine
D000911:Antibodies, Monoclonal; D000939:Epitopes; D019012:Integrin beta1; D021122:Protein Subunits; D011994:Recombinant Proteins