Characterization of a recombinant humanized anti-cocaine monoclonal antibody produced from multiple clones for the selection of a master cell bank candidate.
Characterization of a recombinant humanized anti-cocaine monoclonal antibody produced from multiple clones for the selection of a master cell bank candidate.
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DOI：
10.1016/j.bbrc.2017.04.117
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发表时间：
2017-06-03
影响因子：
3.1
通讯作者：
Norman AB
中科院分区：
文献类型：
--
作者：
Wetzel HN;Webster RP;Saeed FO;Kirley TL;Ball WJ;Norman AB
我们已经产生了人源化的抗可卡因单抗(MAb)，该抗体处于临床前开发的高级阶段。我们在此报告了重组单抗的体外结合亲和力研究、体内药代动力学和药效研究。总体目标是从三个产量最高的转基因克隆中分别鉴定重组抗体，并选择一个以建立主细胞库。在单抗药代动力学研究中，小鼠尾静脉注射H_2E_2(120 mg/kg)后，连续28天采血。用酶联免疫吸附试验测定抗体浓度。H_2E_2浓度与时间的关系符合二室药动学模型。为了测试体内的疗效，小鼠被注射h2E2(120 mg/kg iv)，然后在一小时后注射等摩尔剂量的可卡因。注射可卡因5分钟后采血，取脑组织。用LC/MS法定量可卡因浓度，用[~3H]可卡因结合试验测定抗体与可卡因的亲和力。这三种抗体都有很长的消除半衰期，对可卡因的消除半衰期为2-5 nM，并通过将可卡因隔离在血浆中来阻止可卡因进入大脑。药代动力学和放射性配基结合分析支持将产量最高的克隆(85)指定为主细胞库候选。总体而言，重组h2E2显示出良好的结合特性、药代动力学和体内疗效。
We have generated a humanized anti-cocaine monoclonal antibody (mAb), which is at an advanced stage of pre-clinical development. We report here in vitro binding affinity studies, and in vivo pharmacokinetic and efficacy studies of the recombinant mAb. The overall aim was to characterize the recombinant antibody from each of the three highest producing transfected clones and to select one to establish a master cell bank. In mAb pharmacokinetic studies, after injection with h2E2 (120 mg/kg iv) blood was collected from the tail tip of mice over 28 days. Antibody concentrations were quantified using ELISA. The h2E2 concentration as a function of time was fit using a two-compartment pharmacokinetic model. To test in vivo efficacy, mice were injected with h2E2 (120 mg/kg iv), then one hour later injected with an equimolar dose of cocaine. Blood and brain were collected 5 minutes after cocaine administration. Cocaine concentrations were quantified using LC/MS. The affinity of the antibody for cocaine was determined using a [3H] cocaine binding assay. All three antibodies had long elimination half-lives, 2–5 nM Kds for cocaine, and prevented cocaine’s entry into the brain by sequestering it in the plasma. Pharmacokinetic and radio-ligand binding assays supported designation of the highest producing clone (85) as the master cell bank candidate. Overall, the recombinant h2E2 showed favorable binding properties, pharmacokinetics, and in vivo efficacy.
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影响因子：
4.8
作者：
Kirley TL;Norman AB
通讯作者：
Norman AB
影响因子：
--
作者：
Martell, Bridget A.;Orson, Frank M.;Poling, James;Mitchell, Ellen;Rossen, Roger D.;Gardner, Tracie;Kosten, Thomas R.
通讯作者：
Kosten, Thomas R.
影响因子：
2.5
作者：
Lin, SN;Moody, DE;Foltz, RL
通讯作者：
Foltz, RL
影响因子：
7.3
作者：
Paula, S;Tabet, MR;Ball, WJ
通讯作者：
Ball, WJ
影响因子：
3.9
作者：
Norman, Andrew B.;Gooden, Felicia C. T.;Ball, William J.
通讯作者：
Ball, William J.