Site-specific photocoupling of pBpa mutated scFv antibodies for use in affinity proteomics

Forskningsoutput: TidskriftsbidragArtikel i vetenskaplig tidskrift


Recombinant antibody libraries can provide a source of renewable and high-performing binders tailored for use in affinity proteomics. In this context, the process of generating site-specific 1:1 tagging/functionalization and/or orientated surface immobilization of antibodies has, however, proved to be challenging. Hence, novel ways of generating such engineered antibodies for use in affinity proteomics could have a major impact on array performance. In this study, we have further tailored the design of human recombinant scFv antibodies for site-specific photocoupling through the use of an unnatural amino acid (UAA) and the Dock'n'Flash technology. In more detail, we have generated the 2nd generation of scFvs carrying the photoreactive UAA p-benzoyl-l-phenylalanine (pBpa). Based on key properties, such as expression levels, activity, and affinity, a preferred choice of site for pBpa, located in the beginning of the C-terminal affinity-tag, was for the first time pin-pointed. Further, the results showed that pBpa mutated antibody could be site-specifically photocoupled to free and surface immobilized β-cyclodextrin (an affinity ligand to pBpa). This paves the way for use of scFv antibodies, engineered for site-specific photochemical-based tagging, functionalization, and orientated surface immobilization, in affinity proteomics.


Enheter & grupper
Externa organisationer
  • Aalborg University
  • Lund University

Ämnesklassifikation (UKÄ) – OBLIGATORISK

  • Immunologi inom det medicinska området
  • Annan medicinteknik


Sidor (från-till)985-996
Antal sidor12
TidskriftBiochimica et Biophysica Acta - Proteins and Proteomics
Utgåva nummer8
StatusPublished - 2017 aug
Peer review utfördJa

Relaterad forskningsoutput

Mattias Brofelth, 2018 jan 30, Lund: Department of Immunotechnology, Lund University. 147 s.

Forskningsoutput: AvhandlingDoktorsavhandling (sammanläggning)

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