Production of Novel Camelid Anti-CXCL10 Specific Polyclonal Antibodies and Evaluation of Their Bioreactivity

Abstract

CXCL10 chemokine is a member of CXC chemokine family. It is secreted from a variety of cells in response to IFN- and stimulates a range of inflammatory responses via binding to its receptor, CXCR3. CXCL10 has a pivotal role in the pathogenesis of various infectious diseases, cancers, and inflammatory and autoimmune diseases. It has been put forward as a potential biomarker and therapeutic target in diagnosis and treatment of these diseases. In the present study, production of camel heavy chain antibodies (HCAbs) specific for the CXCL10 is reported. In this regard, recombinant CXCL10 was used for immunization of camel and subsequently the CXCL10 HCAbs were obtained. Afterwards, three subclasses of IgG were separated using protein A and protein G affinity columns, characterized with SDS-PAGE and confirmed for specific binding to the CXCL10 using ELISA. These IgG subclasses successfully recognized CXCL10 and a strong and specific reactivity towards CXCL10 were observed. Therefore, the selected HCAbs and their corresponding expression library could be used to develop a recombinant variable domain of these HCAbs (nanobody or VHH) as a new possible strategy for treatment of multiple sclerosis and other autoimmune diseases.