Directed evolution of an affibody molecule for carbonic anhydrase IX using E. coli display and fluorescence-activated cell sorting

QC20240507
Carbonic anhydrase IX (CAIX) is a transmembrane enzyme that plays an important role in pH regulation and cellular homeostasis. Overexpression of CAIX is commonly observed in various solid tumors, making it an attractive target for cancer therapy. Affibody molecules have previously been developed for CAIX and demonstrated potential as tracers for molecular imaging in murine xenografted tumor models. Here, we further develop such affibody molecules by displaying a designed affibody library on the surface of Escherichia coli, followed by isolation of CAIX-binding variants by a combination of magnetic-assisted cell sorting and fluorescence-activated cell sorting. The sortings were successful and screening of candidates after selection revealed several variants with improved properties, most notably an increase in affinity and CAIX cell binding at 37°C and 42°C.

NATURVETENSKAP Biologi Biokemi och molekylärbiologi hsv//swe
NATURAL SCIENCES Biological Sciences Biochemistry and Molecular Biology hsv//eng
Affibody molecule
carbonic anhydrase IX
Escherichia coli surface display
MACS
FACS
directed evolution
Biotechnology
Bioteknologi

Klint, SusanneAffibody Medical AB, Stockholm, Sweden (författare)
Gunneriusson, ElinAffibody Medical AB, Stockholm, Sweden (författare)
Frejd, FredrikAffibody Medical AB, Stockholm, Sweden (författare)
Ståhl, Stefan,1961-KTH,Albanova VinnExcellence Center for Protein Technology, ProNova,Proteinteknologi(Swepub:kth)u1svy8i4 (författare)
Löfblom, JohnKTH,Skolan för teknikvetenskap (SCI),Proteinvetenskap,Proteinteknologi(Swepub:kth)u1z7cgkl (författare)
KTHProteinvetenskap (creator_code:org_t)

Av författaren/redakt...: Parks, Luke; Klint, Susanne; Gunneriusson, El ...; Frejd, Fredrik; Ståhl, Stefan, 1 ...; Löfblom, John

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