Generation and selection of naïve Fab library for parasitic antigen: Anti‐BmSXP antibodies for lymphatic filariasis

Abstract

Phage display has been applied successfully as a tool for the generation of monoclonal antibodies (mAbs). Naive antibody libraries are unique as they are able to overcome several limitations associated with conventional mAb generation methods like the hybridoma technology. Here, we performed an in vitro selection and generation of Fab antibodies against Brugia malayi SXP protein (BmSXP), a recombinant antigen for the detection of lymphatic filariasis. We developed a naïve multi ethnic Fab antibody library with an estimated diversity of 2.99 × 10 . The antibody library was used to screen for mAbs against BmSXP recombinant antigen. Soluble monoclonal Fab antibodies against BmSXP were successfully isolated from the naïve library. The Fab antibodies obtained were expressed and analyzed to show its binding capability. The diversity obtained from a pool of donors from various ethnic groups allowed for a diverse antibody library to be generated. The mAbs obtained were also functional in soluble form, which makes it useful for further downstream applications. We believe that the Fab mAbs are valuable for further studies and could also contribute to improvements in the diagnosis of filariasis.

Recombinant antigens used as diagnostic tools for lymphatic filariasis

“…The increase in the use of tests based on recombinant antigens for LF diagnosis has the potential to solve the stated limitations related to cross-reactivity and low sensitivity. Further technological advances, as reported in some of the most recent studies using phage display and the BmSXP and BmRI antigens [ 27 , 56 , 58 ], have the potential to facilitate the generation of more efficient antibodies that can be used for such new assays. Another possibility is the use of tests based on chimeric proteins which can potentially enhance the capabilities of the antibody capture assays, as has been done for other diseases [ 89 – 91 ].…”

“…The second study generated Fab antibodies against Bm SXP, selected from a Fab antibody library made with RNA from a pool of B cells that were derived from a large number of healthy blood donors from China, India, and Malaysia. Several clones were selected, with some of those leading to the expression of monoclonal antibodies whose binding to Bm SXP was confirmed through ELISA and pull-down assays [ 58 ].…”

Emergence of zoonotic Brugia pahangi parasite in Thailand

“…parasites. Based on detection of specific IgG4 antibodies against BmR1 ( Brugia Rapid) and BmSXP recombinant antigens [ 54 – 59 ], the commercially available antibody tests for brugian filariasis infection include the Brugia Rapid™ test (Reszon Diagnostics International Sdn. Bhd., Selangor, Malaysia) (https://reszonics.com/products/infectious-diseases-diagnosis/lateral-flow-rapid-test/ Brugia -rapid-test/), and the PanLF Rapid™ test (Reszon Diagnostics International Sdn.…”

Diseño, obtención y caracterización de proteínas recombinantes a partir de determinantes antigénicos asociadas a reacciones alérgicas a ß-lactámicos.

“…169 Las bibliotecas naive utilizan genes reordenados de la región variable procedentes de donantes no inmunizados y, en consecuencia, suelen acceder al repertorio de IgM. 170 Uno de los rasgos distintivos de estas bibliotecas es su naturaleza combinatoria, es decir, durante la construcción de la biblioteca se combinan aleatoriamente cadenas pesadas y ligeras. 171 Muchos antígenos terapéuticos son moléculas propias, pero durante la maduración de los linfocitos B se suelen eliminar los anticuerpos autorreactivos expresados en la superficie de los linfocitos B. Por lo general, se considera que, debido al ensamblaje combinatorio y aleatorio de las cadenas pesadas y ligeras, esta tolerancia inmunológica puede eludirse, al menos parcialmente, permitiendo la generación de anticuerpos dirigidos a autoantígenos.…”