Bispecific antibody modification of nicotinic acetylcholine receptors for biosensing

Real-Time Analysis of Immunogen Complex Reaction Kinetics Using Surface Plasmon Resonance

“…Therefore, the local concentration of linker can be viewed as being very large, which will make the residence time for the unlinked state very short. Our previously experimental results show that in the presence of crosslinkers of different types, e.g., mAb 148 to nAChR or the nAChR/(BsAb) 2 /bovine IgG complex, the channels do remain blocked (18). Since these complexes are known to exist in an equilibrium state, these experiments support the premise that the kinetic onrate of the unlinked end is faster than the channel reconformation rate.…”

“…The mAb 148 hybridoma line was provided by Professor J. M. Lindstrom (University of Pennsylvania). According to previously published procedures (18), MAb 148 IgG 2a was purified from supernatant of cell suspension cultures via affinity chromatography and concentrated using Centricon 30 microconcentrator tubes (Amicon Division of W.R. Grace & Co., Beverly, MA). Goat anti-mouse (anti-MS) IgG (FabЈ-specific), which had been previously adsorbed with rat serum to minimize crossreactions with rat IgG, was purchased from Sigma (St. Louis, MO).…”

“…Membrane vesicles enriched in nAChRs from the electric organ of Torpedo californica were prepared according to the procedures described in past work (18).…”

<title>Gated ion channel biosensor: a functioning nanomachine</title>

“…3x10'°molecules/cm2, G] = 1x108 molecules/cm2, k2D = 1x107 cm2molecules1s' for (1) and 1.25x10" cm2molecules1s for (2). The dimer lifetime (l/k' = l.25x102 s) determines the limiting 'roll over' at high analyte concentrations.…”

“…When 0 falls below 2 the response becomes proportional to [Al112. A further limiting condition is the lifetime of the dimeric channel(2). When this dominates the response time becomes analyte independent and is limited by 1Ik1G.…”