Whole-cell biopanning with a synthetic phage display library of nanobodies enabled the recovery of follicle-stimulating hormone receptor inhibitors

Antibodies, Nanobodies, or Aptamers—Which Is Best for Deciphering the Proteomes of Non-Model Species?

“…In terms of selecting nanobody phage libraries, the process is similar to the phage display libraries used to select for antibodies. Interestingly, these nanobody libraries have recently been used to select for cell surface receptors in situ as well as intracellular targets [62,63], which does open up possibilities for selecting nanobodies against unknown targets on particular sub-populations of cells. Additionally, nanobodies have been generated that are cross-reactive to human and murine proteins [64] or all Trypanosoma species [65].…”

“…Selection can be attempted without knowing the precise target if sufficient materials, such as specific cells, are available. For example, nanobody libraries have been used to select for cell surface receptors [63], though in this case the proteins were expressed in cells prior to selection. This can also be performed using aptamer libraries [81] though it is also possible to "fish" for targets on whole cells or organisms [82,83].…”

“…A schematic is provided in Figure 3 to demonstrate the selection process. The protocol used for generating nanobodies to cell surface receptors used a process similar to SELEX whereby the nanobody library was incubated with target cells for iterative rounds and bound species were re-amplified using Escherichia coli rather than using PCR to re-amplify the bound nucleic acid species [63,85]. This process can be expanded on through the use of tissue sections, and both antibodies and aptamers have been generated to rare cells using both frozen [86,87] and paraffin embedded tissue as the antigen substrate [88].…”

Expression Levels of Follicle-Stimulating Hormone Receptor and Implication in Diagnostic and Therapeutic Strategy of Ovarian Cancer

“…The FSHR belongs to the highly conserved subfamily of the G protein-coupled receptor (GPCR) proteins regulating ovarian follicular maturation and spermatogenesis [6]. Structurally, the FSHR contains a large N-terminal extracellular domain, 7-membrane-spanning (transmembrane) domains, and 3 interconnected loops, and an intracellular C-terminal tail (fig.…”

“…There are also no antibodies available to detect FSHR in a routine pathology institute. Phage display may help to overcome this problem [6]. Current research attempts to demonstrate and quantify the presence of FSHR in tumors and determine their implications in diagnosis and therapy, and much research has been directed to discovering new tumor markers and elucidating their role in the carcinogenesis of ovarian cancer [24].…”