TY - JOUR
T1 - Improved stability of multivalent antibodies containing the human collagen XV trimerization domain
AU - Cuesta, Ángel M.
AU - Sánchez-Martín, David
AU - Blanco-Toribio, Ana
AU - Villate, Maider
AU - Enciso-Álvarez, Kelly
AU - Alvarez-Cienfuegos, Ana
AU - Sainz-Pastor, Noelia
AU - Sanz, Laura
AU - Blanco, Francisco J.
AU - Álvarez-Vallina, Luis
N1 - Funding Information:
Repeated experiments provide an estimate for the error in the This work was supported by grants from the Ministerio de mid-point denaturation temperature of trimerbodyXVIII of ±1°C. Ciencia e Innovación (BIO2008-03233), the Comunidad The denaturation curve of the trimerbodyXV has a smaller signal-Autónoma de Madrid (S-BIO-0236-2006) and the European to-noise ratio than the curve of trimerbodyXVIII because the total Union [SUDOE-FEDER (IMMUNONET-SOE1/P1/E014)] ellipticity change at 210 nm is smaller for this molecule. As a conto L.A.V.; and by grant CTQ2011-28680 to F.J.B. and by sequence, the error in the mid-point temperature determination is grant PS09/00227 from the Fondo de Investigación Sanitaria
PY - 2012
Y1 - 2012
N2 - We recently described the in vitro and in vivo properties of an engineered homotrimeric antibody made by fusing the N-terminal trimerization region of collagen XVIII NC1 domain to the C-terminus of a scFv fragment [trimerbody (scFv-NC1)3; 110 kDa]. Here, we demonstrated the utility of the N-terminal trimerization region of collagen XV NC1 domain in the engineering of trivalent antibodies. We constructed several scFv-based trimerbodies containing the human type XV trimerization domain and demonstrated that all the purified trimerbodies were trimeric in solution and exhibited excellent antigen binding capacity. Importantly, type XV trimerbodies demonstrated substantially greater thermal and serum stability and resistance to protease digestion than type XVIII trimerbodies. In summary, the small size, high expression level, solubility and stability of the trimerization domain of type XV collagen make it the ideal choice for engineering homotrimeric antibodies for cancer detection and therapy.
AB - We recently described the in vitro and in vivo properties of an engineered homotrimeric antibody made by fusing the N-terminal trimerization region of collagen XVIII NC1 domain to the C-terminus of a scFv fragment [trimerbody (scFv-NC1)3; 110 kDa]. Here, we demonstrated the utility of the N-terminal trimerization region of collagen XV NC1 domain in the engineering of trivalent antibodies. We constructed several scFv-based trimerbodies containing the human type XV trimerization domain and demonstrated that all the purified trimerbodies were trimeric in solution and exhibited excellent antigen binding capacity. Importantly, type XV trimerbodies demonstrated substantially greater thermal and serum stability and resistance to protease digestion than type XVIII trimerbodies. In summary, the small size, high expression level, solubility and stability of the trimerization domain of type XV collagen make it the ideal choice for engineering homotrimeric antibodies for cancer detection and therapy.
KW - Antibody engineering
KW - Collagen XV
KW - Collagen XVIII
KW - Multivalent antibody
KW - Tumor targeting
UR - http://www.scopus.com/inward/record.url?scp=84858262290&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84858262290&partnerID=8YFLogxK
U2 - 10.4161/mabs.4.2.19140
DO - 10.4161/mabs.4.2.19140
M3 - Article
C2 - 22453098
AN - SCOPUS:84858262290
SN - 1942-0862
VL - 4
SP - 226
EP - 232
JO - mAbs
JF - mAbs
IS - 2
ER -