■广泛中和抗体是预防和治疗人类免疫缺陷病毒/获得性免疫缺陷综合症(HIV/AIDS)的有希望的候选药物。作为抗逆转录病毒疗法(ART)的替代或联合使用。这些单克隆抗体与病毒附着和进入所必需的病毒位点结合,从而抑制进入宿主细胞。然而,单克隆抗体的成本和可用性,尤其是抗体的组合,在HIV-1流行率最高的中低收入国家(LMICs),阻碍了抗HIVbNAb疗法的实施。
■我们已经生产了三种HIV广中和抗体(bNAb),10-1074,本氏烟草瞬时表达系统中的VRC01和3BNC117。评估了特定修饰对增强效力和功效的影响。为了延长半衰期和增加生物利用度,引入M252Y/S254T/T256E(YTE)或M428L/N434S(LS)突变。为了增加抗体依赖性细胞毒性(ADCC),我们使用糖工程植物系表达了每种抗体的去岩藻糖基化版本。
■大多数bNAb及其变体可以以高达47mg/kg的产量表达。表达系统和修饰均不影响bNAb的中和潜力。无岩藻糖基化的bNAb表现出增强的与FcγRIIIa结合并触发ADCC的能力,无论Fc氨基酸突变的存在。最后,我们证明了在植物中表达的Fc修饰变体显示与FcRn的结合增强,与未修饰的对应物相比,这导致了有利的体内药代动力学特征。
烟草植物是抗HIVbNAb的合适表达宿主,具有增加的功效和改善的药代动力学特征。
UNASSIGNED: Broadly neutralising antibodies are promising candidates for preventing and treating Human Immunodeficiency Virus/Acquired Immunodeficiency Syndrome (HIV/AIDS), as an alternative to or in combination with antiretroviral therapy (ART). These mAbs bind to sites on the virus essential for virus attachment and entry, thereby inhibiting entry into the host cell. However, the cost and availability of monoclonal antibodies, especially combinations of antibodies, hampers implementation of anti-HIV
bNAb therapies in low- to middle- income countries (LMICs) where HIV-1 prevalence is highest.
UNASSIGNED: We have produced three HIV broadly neutralizing antibodies (bNAbs), 10-1074, VRC01 and 3BNC117 in the Nicotiana benthamiana transient expression system. The impact of specific modifications to enhance potency and efficacy were assessed. To prolong half-life and increase bioavailability, a M252Y/S254T/T256E (YTE) or M428L/N434S (LS) mutation was introduced. To increase antibody dependent cellular cytotoxicity (ADCC), we expressed an afucosylated version of each antibody using a glycoengineered plant line.
UNASSIGNED: The majority of bNAbs and their variants could be expressed at yields of up to 47 mg/kg. Neither the expression system nor the modifications impacted the neutralization potential of the bNAbs. Afucosylated bNAbs exhibit enhanced ability to bind to FcγRIIIa and trigger ADCC, regardless of the presence of Fc amino acid mutations. Lastly, we demonstrated that Fc-modified variants expressed in plants show enhanced binding to FcRn, which results in a favourable in vivo pharmacokinetic profile compared to their unmodified counterparts.
UNASSIGNED: Tobacco plants are suitable expression hosts for anti-HIV bNAbs with increased efficacy and an improved pharmacokinetic profile.