ICH Harmonised Tripartite Guideline
Having reached Step 4 of the ICH Process at the ICH Steering Committee meeting on 30 November 1995, this guideline is recommended for adoption to the three regulatory parties to ICH
I. Introduction
This document presents guidance regarding the characterisation of the
expression construct for the production of recombinant DNA protein products in eukaryotic and prokaryotic cells. This document is intended to describe the types of information that are considered valuable in assessing the structure of the expression construct used to produce recombinant DNA derived proteins. This document is not intended to cover the whole quality aspect of rDNA derived
medicinal products. The expression construct is defined as the expression vector containing the coding sequence of the recombinant protein. Segments of the expression construct should be analysed using nucleic acid techniques in conjunction with other tests performed on the purified recombinant protein for assuring the
quality and consistency of the final product. Analysis of the expression construct at the nucleic acid level should be considered as part of the overall evaluation of quality, taking into account that this testing only evaluates the coding sequence of a recombinant gene and not the translational fidelity nor other characteristics of the recombinant protein, such as secondary structure, tertiary structure, and post-translational modifications.
II. Rationale for Analysis of the Expression Construct
The purpose of analysing the expression construct is to establish that the correct coding sequence of the product has been incorporated into the host cell and is maintained during culture to the end of production. The genetic sequence of recombinant proteins produced in living cells can undergo mutations that could alter the properties of the protein with potential adverse consequences to patients. No single experimental approach can be expected to detect all possible modifications to a protein. Protein analytical techniques can be used to assess the amino acid sequence of the protein and structural features of the expressed protein due to post-translational modifications such as proteolytic processing, glycosylation, phosphorylation, and acetylation. Data from nucleic acid analysis may be useful since protein analytical methods may not detect all changes in protein structure resulting from mutations in the sequence coding for the
recombinant protein. The relative importance of nucleic acid analysis and protein analysis will vary from product to product. Nucleic acid analysis can be used to verify the coding sequence and the physical
state of the expression construct. The nucleic acid analysis is performed to ensure that the expressed protein will have the correct amino acid sequence but is not intended to detect low levels of variant sequences. Where the production cells have multiple integrated copies of the expression construct, not all of which may be transcriptionally active, examination of the transcription product itself by analysis of mRNA or cDNA may be more appropriate than analysis of genomic
DNA. Analytical approaches that examine a bulk population of nucleic acids, such as those performed on pooled clones or material amplified by the polymerase Genetic Stability 2 chain reaction, may be considered as an alternative to approaches that depend on
selection of individual DNA clones. Other techniques could be considered that allow for rapid and sensitive confirmation of the sequence coding for the recombinant protein in the expression construct.
Source from ICH, want to know more:https://database.ich.org/sites/default/files/Q5B%20Guideline.pdf
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