Anticuerpos quimiosintéticos como herramienta analíticaobtención y desarrollo

Resumo

Nowadays, antibodies are the major source of reactive molecules and therapeutic biotechnology and because of this; great efforts have been made in recent years to improve these molecules. In order to obtain a fully functional antibody, humanization of antibodies is the most promising technique. However, advances in biotechnology have led to the production of human antibodies which have certain advantages, humanized or chimeric, as mentioned in the covering introduction section. Furthermore, the use of antibody fragments as opposed to whole antibodies has made clear the great advantages of using these molecules (lower immunogenicity, etc.). Due to the success of the derivatization of the fragments with various polymers (i.e. polyethylene glycol (PEG)) this techniques are the most promising prospects. Most of the reports on work carried out to obtain PEGylated fragments have focused on the study of variations in-vivo (pharmacokinetic), without paying attention to important points such as sample preparation and a detailed study of the parameters necessary to achieve a conjugation reaction of great effectiveness. Moreover, very few of them use human antibodies. Therefore, the objective of the work will focus on optimizing the tools necessary to carry out human IgG1 antibody fragmentation to obtain a model adjustable to other fully human antibodies whose therapeutic use is approved as an inhibitor of TNF¿. These fragments are subjected to various PEG conjugations in order to verify the benefits of developing a detailed study with the fragments in order to have a valuable tool in the future for therapeutic use. Fragment and conjugate characterization was performed using enzymatic techniques for both biochemical and immunochemical methods.