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P1.02 Production of polyclonals antibodies against gardnerella vaginalis
  1. Ana Jazmín Otero Cervantes,
  2. Cynthia Rodríguez-Nava,
  3. Karen Cortés Sarabia,
  4. Luz Del Carmen Alarcón Romero,
  5. Isela Parra Rojas,
  6. Amalia Vences Velazquez
  1. Autonomous University of Guerrero, Chilpancingo, Mexico

Abstract

Introduction Bacterial Vaginosis (BV) is the most frequent vaginal infection. It is characterised by a decrease in the number of Lactobacilli and an increse of anaerobic bacteria. Gardnerella vaginalis is the main etiological agent, this bacteria has multiple virulence factors such as the production of biofilm, sialidase and vaginolysin, which can cause the degradation of cervical mucus, adhesion and lysis of epithelial cells. The production of antibodies against this microorganism will allow understanding their role in the development of BV.

Methods Three New Zealand rabbits were immunised for 8 weeks using as antigen the strain ATCC 14018 of G. vaginalis and the complete and incomplete Freund’s adjuvants. The immune response was evaluated at weeks 0, 4th and 7th by indirect ELISA. At the 8th week the rabbits were sacrificed and blood serum was obtained, purification was performed using the Protein A antibody purification kit (Sigma). For the characterisation of the policlonal antibody we perform Indirect ELISA, Dot Blot, Western Blot and inhibition of haemagglutination.

Results Two polyclonal antibodies agains G. vaginalis were obtained. The first was obtained from Rabbit 1 (A.ka. Gv1) and the second one is a Pool (Gv2) from the serum of rabbits 2 and 3. Both antibodies recognise the strain ATCC 14018 of G. vaginalis at titers greater than 1: 2000 and proteins with molecular weights of approximately 38, 50, 65, 75 and 90 kDa, in addition the antibodies are capable of inhibiting lysis of vaginolysin.

Conclusion The produced antibodies will be use to study the pathogenesis of Gardnerella vaginalis during the development of BV.

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