Biopharma Drug Discovery Workflows
with Cells-to-C_T Kits

The precision, sensitivity, and quantitative capabilities of reverse transcription quantitative PCR (RT-qPCR) make it an indispensable technique in the biopharmaceutical industry, helping ensure that products are safe, effective, and of high quality. RT-qPCR serves as a critical pillar of biopharmaceutical development. From biomarker discovery to candidate selection, this highly sensitive and reliable tool enables quantitative genomic analysis, for a wide range of applications.

Within the RT-qPCR process, Invitrogen Cells-to-C_T technology allows the reverse transcription and subsequent qPCR analysis to be performed directly from cell lysates, without the need for RNA purification prior to amplification. This facilitates significant improvements in gene expression analysis workflows which can ultimately lead to increased efficiency.

• Reduced processing steps—eliminates the need for traditional RNA isolation, reducing the number of steps and complexity in the workflow
• Time efficiency—increases efficiency by bypassing RNA purification, allowing fast progression to RT-qPCR
• Minimized sample loss—directly processes cells into cDNA, minimizing sample loss that can occur during RNA extraction procedures
• User-friendly—simplified protocol that is easy to follow, even for those with limited experience in RNA handling

Overview of the TaqMan Cells-to-C_T HepatoExpress workflow. 10 minutes sample preparation (cell lysis) time. Sample to answer in less than 90 minutes. Full workflow steps include:

1. Washing cells and adding lysis solution
2. Transferring lysates to RT plate
3. Reverse transcription
4. Transferring cDNA to qPCR plate
5. qPCR

• Reduced reagent and plastic use—decreases the need for RNA isolation reagents and plastic consumables, which can help lower overall experimental costs and reduce plastic waste
• Less hazardous waste—does not use ethanol, 2-mercaptoehtanol, guanidine thiocyanate, guanidine hydrochloride, or other chaotropic salts for RNA preparation

Comparison of plastic waste generated. Plastic waste generated using (A) the Cells-to-C_T Express kit vs. (B) a traditional, column-based RNA extraction method.

• Scalability—easily scalable to accommodate varying sample sizes, from small studies to large clinical trials
• High throughput capability—facilitates high-throughput processing of multiple samples, which is beneficial for large-scale studies
• Rapid processing—rapid processing of cell lysates helps preserve RNA integrity, reducing degradation risks

Comparison of average C_T values obtained from manual and automated methods. Bars represent the mean ± standard deviation of 192 replicates. Automated and manual methods show nearly identical CT values for both ACTV and CDK4 genes.

Learn more about automating your RT-qPCR workflow.

Application Note:TaqMan Cells-to-C_T Express Kit: automation-friendly product for high-throughput gene expression analysis

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Given its ease of use and reliability, the benefits of RT-qPCR can be applied across the development pipelines of nearly every biopharma research modality. Some examples include:

• Gene expression analysis—analyzing the expression of genes involved in drug targets, pathways, and mechanisms of action. This helps in understanding how drugs affect biological systems.
• Copy number variation (CNV) analysis—assessing copy number variations in cell lines used for biologic production to help ensure genetic stability and consistency.
• SNP genotyping—enables the detection and quantification of specific single nucleotide polymorphisms (SNPs) in a DNA sequence. Researchers can identify specific genetic variations that may confer susceptibility to a particular disease or may affect drug targets, drug metabolism pathways, or drug transport mechanisms.
• Viral titer and vector quantification—quantifying viral vectors used in gene therapy to help ensure the correct dosage and consistency in production.
• Residual DNA testing—detecting and quantifying residual host cell DNA in biologic products to help ensure product safety and compliance with regulatory standards.
• Pathogen detection and adventitious agent testing—screening for microbial contamination in cell cultures, raw materials, and final products. This is crucial for helping maintain the sterility and safety of biopharmaceuticals.
• Biomarker identification and validation—identifying and validating genetic biomarkers that can predict drug response, efficacy, and safety which can support the research and development of personalized medicine.
• Pharmacokinetics and pharmacodynamics (PK/PD) studies—measuring drug levels and their biological effects over time in drug development testing studies to understand the pharmacokinetics and pharmacodynamics of new drugs. Specific focus is often given to the absorption, distribution, metabolism, and excretion (ADME) studies. These studies help provide important information about the drug's behavior in the body and are crucial for understanding its efficacy and safety.
• Stability studies—monitoring the genetic stability of cell banks and final products over time to help ensure long-term product consistency and reliability.