Novel approach for immunoassay capture antibody immobilization – Click chemistry with expanded genetic code
Pihlaja, Riku (2023-11-02)
Novel approach for immunoassay capture antibody immobilization – Click chemistry with expanded genetic code
(02.11.2023)
Julkaisu on tekijänoikeussäännösten alainen. Teosta voi lukea ja tulostaa henkilökohtaista käyttöä varten. Käyttö kaupallisiin tarkoituksiin on kielletty.
avoin
Julkaisun pysyvä osoite on:
https://urn.fi/URN:NBN:fi-fe20231103142909
https://urn.fi/URN:NBN:fi-fe20231103142909
Tiivistelmä
Antibodies are commonly used in biosensors for diagnostics and research due to their ability to specifically
bind certain molecules. However, current manufacturing techniques leave biosensors to use randomly
oriented capture antibodies. This results in decreased antibody function as some antibodies denature,
capture sites are blocked or antibodies wear off. Site specific immobilization has been studied before, but
not with expanded genetic code.
In this study E.coli C.321.deltaA.exp -strain was used to produce fragmented antibodies (Fab) with noncanonical p-azido-l-phenylalanine amino acid incorporated in protein. These fabs were immobilized
covalently in DBCO magnetic beads via bio-orthogonal azide-alkyne cycloaddition reaction. Coated beads
were compared to passively and randomly coated beads by saturating them with labeled antigen.
Results show that Fabs can be modified by adding p-azido-l-phenylalanine into C-terminal end of protein
chain without any compromises to antibody functionality or stability.
bind certain molecules. However, current manufacturing techniques leave biosensors to use randomly
oriented capture antibodies. This results in decreased antibody function as some antibodies denature,
capture sites are blocked or antibodies wear off. Site specific immobilization has been studied before, but
not with expanded genetic code.
In this study E.coli C.321.deltaA.exp -strain was used to produce fragmented antibodies (Fab) with noncanonical p-azido-l-phenylalanine amino acid incorporated in protein. These fabs were immobilized
covalently in DBCO magnetic beads via bio-orthogonal azide-alkyne cycloaddition reaction. Coated beads
were compared to passively and randomly coated beads by saturating them with labeled antigen.
Results show that Fabs can be modified by adding p-azido-l-phenylalanine into C-terminal end of protein
chain without any compromises to antibody functionality or stability.