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In vitro affinity optimization of an anti-BDNF monoclonal antibody translates to improved potency in targeting chronic pain states in vivo.
mAbs ( IF 5.6 ) Pub Date : 2020-04-24 , DOI: 10.1080/19420862.2020.1755000
Edwina Stack 1 , Sheridan McMurray 2 , Gordon McMurray 2 , Jason Wade 1, 3 , Melissa Clark 2 , Gareth Young 2 , Kim Marquette 3 , Sadhana Jain 3 , Kerry Kelleher 3 , Ting Chen 3 , Qingcong Lin 3 , Laird Bloom 3 , Laura Lin 3 , William Finlay 1 , Rie Suzuki 2 , Orla Cunningham 1
Affiliation  

The role of brain-derived neurotrophic factor (BDNF) signaling in chronic pain has been well documented. Given the important central role of BDNF in long term plasticity and memory, we sought to engineer a high affinity, peripherally-restricted monoclonal antibody against BDNF to modulate pain. BDNF shares 100% sequence homology across human and rodents; thus, we selected chickens as an alternative immune host for initial antibody generation. Here, we describe the affinity optimization of complementarity-determining region-grafted, chicken-derived R3bH01, an anti-BDNF antibody specifically blocking the TrkB receptor interaction. Antibody optimization led to the identification of B30, which has a > 300-fold improvement in affinity based on BIAcore, an 800-fold improvement in potency in a cell-based pERK assay and demonstrates exquisite selectivity over related neurotrophins. Affinity improvements measured in vitro translated to in vivo pharmacological activity, with B30 demonstrating a 30-fold improvement in potency over parental R3bH01 in a peripheral nerve injury model. We further demonstrate that peripheral BDNF plays a role in maintaining the plasticity of sensory neurons following nerve damage, with B30 reversing neuron hyperexcitability associated with heat and mechanical stimuli in a dose-dependent fashion. In summary, our data demonstrate that effective sequestration of BDNF via a high affinity neutralizing antibody has potential utility in modulating the pathophysiological mechanisms that drive chronic pain states.

中文翻译:

抗BDNF单克隆抗体的体外亲和力优化可提高体内靶向慢性疼痛状态的能力。

脑源性神经营养因子(BDNF)信号传导在慢性疼痛中的作用已得到充分证明。鉴于BDNF在长期可塑性和记忆力中的重要中心作用,我们试图设计一种针对BDNF的高亲和力,受周边限制的单克隆抗体来调节疼痛。BDNF在人类和啮齿动物之间具有100%的序列同源性;因此,我们选择了鸡作为初始抗体生成的替代免疫宿主。在这里,我们描述了互补决定区移植的,鸡源性的R3bH01的亲和力优化,R3bH01是一种特异性阻断TrkB受体相互作用的抗BDNF抗体。抗体的优化导致了B30的鉴定,基于BIAcore,它的亲和力提高了300倍以上,在基于细胞的pERK分析中,效价提高了800倍,并证明了相对于相关神经营养蛋白的精湛选择性。体外测量的亲和力改善转化为体内药理活性,在周围神经损伤模型中,B30比亲本R3bH01的效力提高了30倍。我们进一步证明,外周BDNF在维持神经损伤后感觉神经元的可塑性方面发挥作用,B30以剂量依赖的方式逆转与热和机械刺激相关的神经元过度兴奋。总之,我们的数据表明,通过高亲和力中和抗体有效隔离BDNF在调节驱动慢性疼痛状态的病理生理机制方面具有潜在的实用性。体外测量的亲和力改善转化为体内药理活性,在周围神经损伤模型中,B30比亲本R3bH01的效力提高了30倍。我们进一步证明,外周BDNF在维持神经损伤后感觉神经元的可塑性方面发挥作用,B30以剂量依赖性方式逆转与热和机械刺激有关的神经元过度兴奋。总之,我们的数据表明,通过高亲和力中和抗体有效隔离BDNF在调节驱动慢性疼痛状态的病理生理机制方面具有潜在的实用性。体外测量的亲和力改善转化为体内药理活性,在周围神经损伤模型中,B30比亲本R3bH01的效力提高了30倍。我们进一步证明,外周BDNF在维持神经损伤后感觉神经元的可塑性方面发挥作用,B30以剂量依赖性方式逆转与热和机械刺激有关的神经元过度兴奋。总而言之,我们的数据表明通过高亲和力中和抗体有效隔离BDNF在调节驱动慢性疼痛状态的病理生理机制方面具有潜在的实用性。我们进一步证明,外周BDNF在维持神经损伤后感觉神经元的可塑性方面发挥作用,B30以剂量依赖性方式逆转与热和机械刺激有关的神经元过度兴奋。总之,我们的数据表明,通过高亲和力中和抗体有效隔离BDNF在调节驱动慢性疼痛状态的病理生理机制方面具有潜在的实用性。我们进一步证明,外周BDNF在维持神经损伤后感觉神经元的可塑性方面发挥作用,B30以剂量依赖的方式逆转与热和机械刺激相关的神经元过度兴奋。总而言之,我们的数据表明通过高亲和力中和抗体有效隔离BDNF在调节驱动慢性疼痛状态的病理生理机制方面具有潜在的实用性。
更新日期:2020-04-24
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