Multicolor chromosome bar codes

Müller, Stefan; Wienberg, Johannes

doi:10.1159/000094208

Cited by 9 publications

(3 citation statements)

References 52 publications

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“…Rx-FISH and somatic cell hybrid-based CBC are reviewed in another paper of this special issue (Müller and Wienberg, 2006); thus, this approach is only described here in short:…”

Section: Fish-banding Methods In Humanmentioning

confidence: 99%

Multicolor fluorescence in situ hybridization (FISH) applied to FISH-banding

Liehr

Starke

Heller

et al. 2006

Cytogenet Genome Res

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During the last decade not only multicolor fluorescence in situ hybridization (FISH) using whole chromosome paints as probes, but also numerous chromosome banding techniques based on FISH have been developed for the human and for the murine genome. This review focuses on such FISH-banding techniques, which were recently defined as ‘any kind of FISH technique, which provide the possibility to characterize simultaneously several chromosomal subregions smaller than a chromosome arm. FISH-banding methods fitting that definition may have quite different characteristics, but share the ability to produce a DNA-specific chromosomal banding’. While the standard chromosome banding techniques like GTG lead to a protein-related black and white banding pattern, FISH-banding techniques are DNA-specific, more colorful and, thus, more informative. For some, even high-resolution FISH-banding techniques the development is complete and they can be used for whole genome hybridizations in one step. Other FISH-banding methods are only available for selected chromosomes and/or are still under development. FISH-banding methods have successfully been applied in research in evolution- and radiation-biology, as well as in studies on the nuclear architecture. Moreover, their suitability for diagnostic purposes has been proven in prenatal, postnatal and tumor cytogenetics, indicating that they are an important tool with the potential to partly replace the conventional banding techniques in the future.

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“…Rx-FISH and somatic cell hybrid-based CBC are reviewed in another paper of this special issue (Müller and Wienberg, 2006); thus, this approach is only described here in short:…”

Section: Fish-banding Methods In Humanmentioning

confidence: 99%

Multicolor fluorescence in situ hybridization (FISH) applied to FISH-banding

Liehr

Starke

Heller

et al. 2006

Cytogenet Genome Res

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“…The procedure and imaging was performed by ChromBios GmbH and described in reference. 38 In brief, about 2 μg of plasmid DNA were labeled with Digoxigenin (dig-dUTP, Roche) by standard nick translation procedures. About 40 ng were used per 10 μl hybridization mixture.…”

Section: Fluorescence In-situ Hybridizationmentioning

confidence: 99%

Retrospective assessment of clonal origin of cell lines

Langsdorf

Kanevskaia

et al. 2021

Biotechnol Progress

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Cell lines used for the manufacture of recombinant proteins are expected to arise from a single cell as a control strategy to limit variability and ensure consistent protein production. Health authorities require a minimum of two rounds of limiting dilution cloning or its equivalent to meet the requirement of single cell origin. However, many legacy cell lines may not have been generated with process meeting this criteria potentially impeding the path to commercialization. A general monoclonality assessment strategy was developed based on using the site of plasmid integration for a cell's identity. By comparing the identities of subclones from a master cell bank (MCB) to each other and that of the MCB, a probability of monoclonality was established. Two technologies were used for cell identity, Southern blot and a PCR assay based on plasmid-genome junction sequences identified by splinkerette PCR.Southern blot analysis revealed that subclones may have banding patterns that differ from each other and yet indicate monoclonal origin. Splinkerette PCR identifies cellular sequence flanking the point(s) of plasmid integration. The two assays together provide complimentary data for cell identity that enables proper monoclonality assessment and establishes that the three legacy cell lines investigated are all of clonal origin.

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“…If a consistent FISH pattern is observed in analyzed metaphase cells, the cell line can be deemed of clonal origin. The same slide can also be counterstained using multicolor-FISH (M-FISH) (20). M-FISH consists of "painting" the arrested metaphase chromosomes with a specific mixture of fluorochrome probes for the different Chinese hamster chromosomes.…”

Section: Figurementioning

confidence: 99%

Advancing Biologics Development Programs with Legacy Cell Lines: Advantages and Limitations of Genetic Testing for Addressing Clonality Concerns Prior to Availability of Late Stage Process and Product Consistency Data

Hartman

Almond

et al. 2019

PDA Journal of Pharmaceutical Science and Technology

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The bioprocessing industry uses recombinant mammalian cell lines to generate therapeutic biologic drugs. To ensure consistent product quality of the therapeutic proteins, it is imperative to have a controlled production process. Regulatory agencies and the biotechnology industry consider cell line "clonal origin" an important aspect of maintaining process control. Demonstration of clonal origin of the cell substrate, or production cell line, has received considerable attention in the past few years, and the industry has improved methods and devised standards to increase the probability and/or assurance of clonal derivation. However, older production cell lines developed before the implementation of these methods, herein referred to as "legacy cell lines," may not meet current regulatory expectations for demonstration of clonal derivation. In this article, the members of the IQ Consortium Working Group on Clonality present our position that the demonstration of process consistency and product comparability of critical quality attributes throughout the development life cycle should be sufficient to approve a license application without additional genetic analysis to support clonal origin, even for legacy cell lines that may not meet current day clonal derivation standards. With this commentary, we discuss advantages and limitations of genetic testing methods to support clonal derivation of legacy cell lines and wish to promote a mutual understanding with the regulatory authorities regarding their optional use during early drug development, subsequent to Investigational New Drug (IND) application and before demonstration of product and process consistency at Biologics License Applications (BLA) submission.

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Multicolor chromosome bar codes

Cited by 9 publications

References 52 publications

Multicolor fluorescence in situ hybridization (FISH) applied to FISH-banding

Multicolor fluorescence in situ hybridization (FISH) applied to FISH-banding

Retrospective assessment of clonal origin of cell lines

Advancing Biologics Development Programs with Legacy Cell Lines: Advantages and Limitations of Genetic Testing for Addressing Clonality Concerns Prior to Availability of Late Stage Process and Product Consistency Data

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