Journal
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
Volume 23, Issue 15, Pages -Publisher
MDPI
DOI: 10.3390/ijms23158470
Keywords
biopanning; phage display; mouse common beta receptor; monoclonal antibodies; functional inhibition
Funding
- Australian Research Council Industrial Transformation Training Centres (ARC ITTC) under the ARC Training Centre for Biopharmaceutical Innovation (CBI) [IC160100027]
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Monoclonal antibodies (mAbs) are highly successful and versatile protein-based pharmaceutical products used in the treatment of various pathological conditions. The quality and presentation format of the antigen play a crucial role in the selection outcomes of mAbs against membrane protein targets. Soluble presentation of a type I membrane protein is more advantageous in isolating functional mAbs with higher sequence diversity.
Monoclonal antibodies (mAbs) are one of the most successful and versatile protein-based pharmaceutical products used to treat multiple pathological conditions. The remarkable specificity of mAbs and their affinity for biological targets has led to the implementation of mAbs in the therapeutic regime of oncogenic, chronic inflammatory, cardiovascular, and infectious diseases. Thus, the discovery of novel mAbs with defined functional activities is of crucial importance to expand our ability to address current and future clinical challenges. In vitro, antigen-driven affinity selection employing phage display biopanning is a commonly used technique to isolate mAbs. The success of biopanning is dependent on the quality and the presentation format of the antigen, which is critical when isolating mAbs against membrane protein targets. Here, we provide a comprehensive investigation of two established panning strategies, surface-tethering of a recombinant extracellular domain and cell-based biopanning, to examine the impact of antigen presentation on selection outcomes with regards to the isolation of positive mAbs with functional potential against a proof-of-concept type I cell surface receptor. Based on the higher sequence diversity of the resulting antibody repertoire, presentation of a type I membrane protein in soluble form was more advantageous over presentation in cell-based format. Our results will contribute to inform and guide future antibody discovery campaigns against cell surface proteins.
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