TATAA Blog

Comparing qPCR vs PCR

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qPCR has emerged as an essential tool for quantitative assessments like biodistribution, shedding, pharmacokinetics and transgene expression. TATAA biocenter specializes in assay development, optimization, qualification, validation and analysis of qPCR and dPCR.

Applications for qPCR vs PCR

Cell and gene therapy, with their potential to cure a range of diseases, have emerged as promising medical avenues. In this context, Polymerase Chain Reaction (PCR) and quantitative Polymerase Chain Reaction (qPCR) are not just tools but indispensable pillars of drug development. They play a pivotal role in assessing the efficacy and safety of cell and gene therapy products. Both PCR and qPCR are amplifications of targeted gene sequences, but the key difference is that qPCR quantifies the target, whereas PCR does not.

Principles of PCR and qPCR

PCR is a molecular biology technique used to amplify a specific segment of DNA through a series of temperature-controlled cycles. It involves three main steps: denaturation, annealing, and extension, facilitated by a DNA polymerase enzyme. In contrast, qPCR, also known as real-time PCR, allows for quantifying DNA or RNA molecules in a sample. It measures the accumulation of PCR products in real-time using fluorescent probes or dyes, providing quantitative data throughout the amplification process.

Applications in Cell and Gene Therapy Drug Development

PCR and qPCR find extensive applications in cell and gene therapy drug development. In preclinical studies, PCR techniques are used to characterize vectors, while qPCR is employed to verify transgene expression and assess the purity of cell populations. During clinical trials, qPCR is instrumental in monitoring patient response, detecting potential adverse events, and ensuring the stability of therapeutic products. Furthermore, qPCR is indispensable in the quality control of final drug products, where it assesses vector integrity, confirms transgene expression levels, and detects contaminants.

Advantages of qPCR over PCR

Quantitative PCR offers several advantages over conventional PCR in cell and gene therapy drug development. Firstly, qPCR provides quantitative data, allowing for precise measurement of nucleic acid concentrations. This quantitative information is essential for determining transgene expression levels, biodistribution, and pharmacokinetics. Secondly, qPCR is very sensitive, enabling the detection of low-copy number targets with higher accuracy. Additionally, qPCR enables high-throughput analysis through automation, improving efficiency in large-scale studies.

Differences between qPCR and PCR

Guide on PCR in drug development

Download PCR guide from qPCR VS PCR blog post.

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Limitations and Challenges

Despite its advantages, qPCR also has limitations and challenges in cell and gene therapy drug development. One limitation is the potential for PCR inhibitors in complex biological samples, which can affect quantification accuracy. It is, therefore, essential to include controls to monitor potential inhibitory effects. Digital PCR can be used instead for extra challenging matrices like shedding matrices since digital PCR is less sensitive to inhibitory agents. Moreover, designing specific and efficient primers and probes for qPCR assays can be difficult, particularly for complex target sequences with highly homologous sequences. We highly recommend outsourcing your assay design and validation to a laboratory like ours, which has more than two decades of experience in qPCR, to achieve a fast and reliable outcome.

Emerging Trends and Future Potential

The advancements in PCR and qPCR technologies drive innovation and shape the future of cell and gene therapy drug development. The integration of digital PCR (dPCR) techniques, for instance, offers enhanced sensitivity and absolute quantification capabilities. The development of multiplex qPCR assays allows for the simultaneous detection and quantification of multiple targets, thereby improving efficiency and reducing sample volume, assay time, and cost. The adoption of novel detection chemistries and platforms is enabling more robust and reliable quantification of nucleic acids in complex samples. 

qPCR for Protein Quantification Using Olink

Olink provides high-multiplex protein biomarker analysis solutions utilizing antibodies for specificity and oligo barcodes quantified with qPCR for sensitivity. The assays enable the simultaneous measurement of up to 3,072 proteins from small sample volumes. Olink offers protein panels with flexible options for targeted and discovery-based protein biomarker research. Their extensive panel library covers various biological processes, including immunology, oncology, and neurology, empowering researchers to gain deep insights into disease mechanisms and identify potential biomarkers.

TATAA Biocenter has been a certified Olink service provider for many years, possessing extensive knowledge about the different platforms and panels.

qPCR Service Provider for Cell and Gene therapies

TATAA Biocenter is a Contract Research Organization (CRO) specializing in qPCR and dPCR assays for biomarker profiling and bioanalysis. We offer assay development, optimization, qualification, validation, and high-throughput analysis from extraction to data transfer. Our team of experts in nucleic acid quantification operates from a purpose-built laboratory equipped with the latest instrumentation and infrastructure. Contact us for a consultation on your qPCR needs.

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