Immunization of alpaca and camel with customized antigens to achieve ideal titers
Phage display nanobody library construction with diversity >109 cfu
Synthetic nanobody library (>1010 cfu) to bypass the immunization
Library selection by both in vitro (solid-phase, solution-phase, cell-based) panning and in vivo panning technology
Primary positive clone identification by ELISA and subsequential Sanger DNA sequencing and/or next-generation sequencing
High-throughput nanobody expression in either E. coli cells or mammalian cells with hexa His or Fc-tag
Binding confirmation by ELISA/FACS and quantified by BLI and/or SPR
In vitro function assays, internalization, neutralization, activation, proliferation, ADCC, etc.
Computer-assisted and/or structure-driven nanobody engineering, humanization, affinity maturation (up to 1,000 fold), etc.
High-throughput nanobody stability evaluation by Uncle (Tm, Tagg)
Multiple chromatography to evaluate nanobody aggregation, hydrophobicity, cross-reactivity, etc.
Nanobody derivates, ADC, ARC, Bi-specific, CAR, etc.
Uncle
BTX Electroporation System
ForteBio Octet 384
BIACore 8K
BECKMAN Cytoflex
PerkinElemer EnVision
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