在knob-into-hole技术的基础上,单特异性抗体分子上加了Fc-dummy chains,避免半抗体样分子(half-antibody like molecules)聚集,形成二聚体。在input A和Input B加入的两个半抗体样分子相遇,则发生Format Chain交换,两个Fc-dummy chains带有不同电荷,及Konb...
在knob-into-hole技术的基础上,单特异性抗体分子上加了Fc-dummy chains,避免半抗体样分子(half-antibody like molecules)聚集,形成二聚体。在input A和Input B加入的两个半抗体样分子相遇,则发生Format Chain交换,两个Fc-dummy chains带有不同电荷,及Konb和hole,自组装成二聚体(Dummy dimmer)在纯化时,可以被去掉。
However, the bsAb byproducts and impurities, including mispaired homodimers, half-antibodies, light chain mispairings, antibody fragments and high levels of high molecular weight (HMW) species, all pose unique challenges to their downstream processing. Here, using two knob-into-hole (KiH) ...
bispecific antibodyknob-into-holechromatography purificationlow pH washhole-hole homodimeraggregation propensityBispecific antibodies (bsAbs) are therapeutically promising due to their ability to bind to two different antigens. However, the bsAb byproducts and impuritiesZhang, Wei...
bispecific antibodyassemblypurificationknobs into holeshigh‐throughput screeningProduction of knob and hole dual light chain bispecific antibodies poses several unique challenges for development of a feasible industrial scale manufacturing process. We developed an efficient process for the assembly and ...
Antiparallel Conformation of Knob and Hole Aglycosylated Half-Antibody Homodimers Is Mediated by a C H 2-C H 3 Hydrophobic Interaction. J Mol Biol 2014 ; 426(9): 1947-57; PMID: 24576605 ; http://dx.doi.org/ 10.1016/j.jmb.2014.02.015...
Bispecific antibodyKnob-into-holeFlow-through polishingHost cell proteinsHigh molecular weight impuritiesLow molecular weight impuritiesHigher aggregation propensityBispecific antibodies (bsAbs), though possessing great therapeutic potential, are extremely challenging to obtain at high purity within a limited ...
The Knob-into-Hole (KiH) technology uses complementary mutations in the CH3 region of the antibody Fc fragment to achieve heavy chain heterodimerization. Here we describe the X-ray crystal structures of glycosylated and disulfide-engineered heterodimeric KiH Fc fragment and its homodimeric Knob-Knob ...
In addition, the inevitable expression of homodimers in host cells presents further obstacles to the commercial development of bispecific antibodies as therapeutics. These byproducts, which share similar physicochemical properties with the target, pose several challenges for downstream purification processes....