PDA TR14_柱层分离工艺_Column Separation Process_1992

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2024年6月2日发(作者：)

Industry Perspective on the Validation of Column-Based

Separation Processes for the Purification of Proteins

蛋白质净化柱层分离工艺验证的工业观点

I. Foreword 前言

The purpose of this document is to outline some of the significant issues related to

the validation of column-based separation processes used in the purification of

proteins produced by recombinant DNA (rDNA), hybri-doma technology, or peptide

synthesis. While validation of these processes has been identified as a first priority by

the PDA Biotechnology Task Force, the issues raised in certain sections of this

document may have broader applications, including processes for non-protein phar-

maceutical products purified by HPLC, as well as protein pharmaceuticals which are

not produced by rDNA, hybridoma, or peptide synthesis technologies.

此文件目的是列出与DNA重组 (rDNA)，杂交瘤技术或肽合成产生的蛋白质净化所使用

的柱层分离工艺验证相关的一些重要问题。这些工艺的验证是PDA生物技

术特别小组最重要的任务，此文件某些章节提到的问题可能有更广泛的应用，包括由

HPLC净化的非蛋白质药品工艺，非DNA重组 (rDNA)，杂交瘤技术或肽合成技术产生的

蛋白质产品。

While column-based separations are key purification techniques in the production

of recombinant proteins and monoclonal antibodies, little has been written regarding

the validation of these processes. In general, process validation is the assurance that

product quality is derived from careful attention to a number of factors, including

process design, selection and use of quality parts and materials, and control of the

process through appropriate in-process and end-product testing. In May 1987, the

Center for Drugs and Biologies and the Center for Devices and Radiological Health of

the Food and Drug Administration published the "Guideline on General Principles of

Process Validation." While this guideline is useful, it does not include all of the specific

elements required in the validation of a manufacturing process. Therefore, the PDA

established the Biotechnology Task Force on Purification and Scale-up to develop a

practical guide for the validation of column-based separations used in the manufacture

of proteins intended for use as therapeutic or diagnostic agents. 柱层分离是重组蛋白

质和单克隆抗体类生产的关键净化技术，这些工艺验证文件很少。一般来说，工艺验证

是为确保产品质量经过谨慎处理，包括工艺设计，选择，质量部分和材料的使用，适当

的过程中测试及制成品测试的工艺控制等因素。1987年5月，药品和生物中心，设备中心

和食品药品管理局放射卫生组出版了“工艺验证一般原则指南”。虽然这个指南有其用

途，但不包括生产工艺验证要求的所有指定部件。因此，PDA成立了净化和工艺放大生

物技术特别小组，以建立用于治疗剂或诊断剂中蛋白质生产使用的柱层分离验证的实用

指南。

The goal of validation is to demonstrate that a process, when operated within

established limits, produces a product of appropriate and consistent quality. While it

may seem that successful validation involves an inordinate amount of work over a long

period of time and consumes scarce resources, a strategy which provides for the

validation of critical process parameters is vital from both a quality and business

perspective. Because no two processes are identical, it is impossible to define exactly

what work should be done to accomplish a satisfactory validation. During validation,

the critical process parameters should be identified, and, based on sound scientific

principles, appropriate studies should be performed to demonstrate that the parame-

ters can be met on a consistent basis. 验证目的是证明工艺在规定范围内操作会产生

合适且质量相同的产品。成功的验证似乎需要长时间的过量工作，且消耗稀少资源，但

关键工艺参数验证的方法从质量和商业角度都是至关重要的。因为没有两个工艺是相同

的，所以不可能准确定义如何完成满意的验证。验证过程中，应确定关键工艺参数，根

据合理的科学原理进行相关研究，以证明可以持续达到参数要求。

Validation should be considered as early in the development of a process as is

practical. In this way, data required for validation can be collected during development

studies and the production of batches for clinical studies. The most appropriate

validation data are collected during the production of products to be used in Phase III

clinical trials. In addition to the in-process testing, the evaluation of product in humans

under carefully monitored clinical trials provides the ultimate test of the safety and

efficacy of the product. 工艺前期应考虑验证是否实际。从而验证需要的数据可以在开

发研究和临床研究生产过程中进行收集。最重要的验证数据可在临床试验第3阶段的产品

生产过程中收集。除了过程中测试，临床试验人员仔细监测的产品评估将提供产品安全

性和效力的最终测试。

In preparing this document, we have assumed that the reader is familiar with

protein purification in general and column-based separations in particular. For back-

ground reading on this subject, the reader is referred to references (1-5). 文件编制

时，我们假设读者都熟悉蛋白质净化，特别是柱层分离。关于此主题的背景材料，见参

考文件（1-5）。

II. Introduction 介绍

Biotechnology in the pharmaceutical industry combines advances in genetic

engineering, cell fusion technology, classical biochemistry and microbiology, traditional

pharmaceutical technology, and biochemical engineering to produce once scarce

proteins for testing as potential therapeutic or diagnostic agents. These proteins are

produced in a suitable host cell by fermentation or cell culture processes. The protein

may be either retained intracellularly or secreted into the culture medium. If the protein

remains inside the cell, then the cells must be harvested, disrupted, and the debris that

is created removed to yield a particulate-free extract for further purification. If the

protein is secreted, the cells must be separated from the conditioned culture medium

prior to purification. These steps, shown schematically in Figure 1, are often referred to

as upstream processing, isolation, or primary separation steps. These upstream steps

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