Improvement of the methodological strategies to product functionalizes antibodies using Small Angle Neutron Scattering (SANS)

Abstract

INTRODUCTION Antibodies are used by jawed vertebrates for defense against invading pathogens. Usage of those versatile tools in a plethora of settings in clinics and biomedical sciences hinges on functionalization strategies that retain native antibody reactivity. To this date, antibody functionalization is performed by trial and error. OBJECTIVES We aim to reduce costs by providing general principles to allow the full spectrum of antibody functionalization by correlating functionalized antibody reactivity to cognate antigen by small angle neutron scattering, SANS, measurements and mathematical modeling of antibody and antibody-antigen super-complexes, obtained by titration experiments. MATERIALS AND METHODS For this research we have used for as antibody pure goat anti rabbit immunoglobulin, and for the antigen, pure Horseradish Peroxidase Preliminary results show that the systems (antibody and antibody-antigen complexes) do not change in the range of a temperature related to storage temperature (25º C), body temperature (37º C) and 40º C. DISCUSSION AND RESULTS These results will give us the pair distribution function of these systems and the results will be viewed in light of published precedence to highlight areas where future effort is needed to refine such versatile tools and improve their production. However, between the antibody and the complexes structure, different conformations were observed. The antibody has a globular structure with a radius of gyration around 33 Å, and the complexes display an elongated cylindrical shape with radius of gyration around 63 Å. CONCLUSION This study shows how the scattering techniques (SANS) can provide useful information about the conformation of the antibody and antibody-antigen formation and help to shed light in the understanding the physical, chemical, and structural changes on the organization of these important antibody functionalization for the immunological system.