Optimization of the Imaged cIEF Method for Monitoring the Charge Heterogeneity of Antibody-Maytansine Conjugate.

IF 2.3 3区化学 Q3 CHEMISTRY, ANALYTICAL

Journal of Analytical Methods in Chemistry Pub Date : 2023-01-01 DOI:10.1155/2023/8150143

Ayat Abbood

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引用次数: 1

Abstract

The aim of this study was to develop a whole-column imaging-detection capillary isoelectric focusing (icIEF) method for the analytical characterization of charge heterogeneity of a novel humanized anti-EphA2 antibody conjugated to a maytansine derivative. In addition to focusing time, sample composition was optimized: pH range, percent of carrier ampholytes, conjugated antibody concentration, and urea concentration. A good separation of charge isoforms was obtained with 4% carrier ampholytes of a large (3-10) and narrow pH range (8-10.5) (1 : 1 ratio), conjugated antibody concentration (0.3-1 mg/ml) with a good linearity (R²: 0.9905), 2 M of urea concentration, and 12 minute for focusing. The optimized icIEF method demonstrated a good interday repeatability with RSD values: <1% (pI), <8% (% peak area), and 7% (total peak areas). The optimized icIEF was useful as an analytical characterization tool to assess the charged isoform profile of a discovery batch of the studied maytansinoid-antibody conjugate in comparison to its naked antibody. It exhibited a large pI range (7.5-9.0), while its naked antibody showed a narrow pI range (8.9-9.0). In the discovery batch of maytansinoid-antibody conjugate, 2% of charge isoforms had the same pI as the pI of naked antibody isoforms.

查看原文本刊更多论文

检测抗体-美坦辛偶联物电荷异质性的cIEF成像方法优化。

本研究的目的是建立一种全柱成像检测毛细管等电聚焦(icIEF)方法，用于分析一种新型人源抗epha2抗体与美坦辛衍生物偶联的电荷异质性。除了聚焦时间外，还对样品组成进行了优化:pH范围、载体两性水解物百分比、偶联抗体浓度和尿素浓度。采用4%的大(3-10)窄pH范围(8-10.5)(1:1比例)载体两性电解质，偶联抗体浓度(0.3-1 mg/ml)，线性良好(R2: 0.9905)，尿素浓度为2 M，聚焦时间为12 min，获得了良好的电荷异构体分离效果。优化后的icIEF方法具有良好的日间重复性，RSD值为:

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Analytical Methods in Chemistry CHEMISTRY, ANALYTICAL-ENGINEERING, CIVIL

CiteScore

4.80

自引率

3.80%

发文量

审稿时长

6-12 weeks

期刊介绍： Journal of Analytical Methods in Chemistry publishes papers reporting methods and instrumentation for chemical analysis, and their application to real-world problems. Articles may be either practical or theoretical. Subject areas include (but are by no means limited to): Separation Spectroscopy Mass spectrometry Chromatography Analytical Sample Preparation Electrochemical analysis Hyphenated techniques Data processing As well as original research, Journal of Analytical Methods in Chemistry also publishes focused review articles that examine the state of the art, identify emerging trends, and suggest future directions for developing fields.