TY - JOUR
T1 - Study of an anti-human transthyretin immunoadsorbent. Influence of coupling chemistry on binding capacity and ligand leakage
AU - Riedstra, Symon
AU - Ferreira, João Paulo M.
AU - Costa, Paulo M.P.
N1 - Funding Information:
We gratefully acknowledge the generous support of Hospital Geral de Santo António, Oporto. We also would like to thank the Nuclear Medicine Department of lnstituto Português de Oncologia, Oporto, for letting us use their laboratory. This work was supported by grant PBICT/C/1981/95 from Junta Nacional de Investigação Cientı́fica e Tecnológica (JNICT), Ministério da Ciência e Tecnologia, Portugal.
PY - 1998/2/13
Y1 - 1998/2/13
N2 - A variant of transthyretin (TTR Val30Met) has been identified as the main protein precursor of the amyloid fibrils deposited in familial amyloidotic polyneuropathy (FAP). Specific removal of TTR in an extracorporeal immunoadsorption procedure is currently under investigation as a possible treatment of FAP. Immunoadsorbents were constructed by immobilizing murine anti-TTR monoclonal antibody 88.6.BA9 onto agarose gel supports via several different coupling chemistries. The influence of coupling conditions such as pH and antibody density, and of perfusion variables, such as antigen concentration and applied flow-rate, on the TTR capture efficiency, was determined. Cyanogen bromide-, carbonyldiimidazole- and aldehyde-activated (ALD) supports conjugated with antibody at optimal pH, provided immunoadsorbents with comparable TTR binding capacities. Regarding stability, leakage was lowest for the ALD based immunoadsorbents, particularly at high pH.
AB - A variant of transthyretin (TTR Val30Met) has been identified as the main protein precursor of the amyloid fibrils deposited in familial amyloidotic polyneuropathy (FAP). Specific removal of TTR in an extracorporeal immunoadsorption procedure is currently under investigation as a possible treatment of FAP. Immunoadsorbents were constructed by immobilizing murine anti-TTR monoclonal antibody 88.6.BA9 onto agarose gel supports via several different coupling chemistries. The influence of coupling conditions such as pH and antibody density, and of perfusion variables, such as antigen concentration and applied flow-rate, on the TTR capture efficiency, was determined. Cyanogen bromide-, carbonyldiimidazole- and aldehyde-activated (ALD) supports conjugated with antibody at optimal pH, provided immunoadsorbents with comparable TTR binding capacities. Regarding stability, leakage was lowest for the ALD based immunoadsorbents, particularly at high pH.
KW - Familial amyloidotic polyneuropathy
KW - Immobilization
KW - Immunoadsorbents
KW - Monoclonal antibodies
KW - Transthyretin
UR - http://www.scopus.com/inward/record.url?scp=18144436780&partnerID=8YFLogxK
U2 - 10.1016/S0378-4347(97)00546-X
DO - 10.1016/S0378-4347(97)00546-X
M3 - Article
C2 - 9521557
AN - SCOPUS:18144436780
SN - 1572-6495
VL - 705
SP - 213
EP - 222
JO - Journal of Chromatography B: Biomedical Applications
JF - Journal of Chromatography B: Biomedical Applications
IS - 2
ER -