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Analysis Infectivity and Titer Determination of Viruses and Viral vectors of Biopharmaceutical Compounds by Quantitative Polymerase Chain Reaction (qPCR) (CAT#: STEM-B-0386-CJ)

Introduction

Biological activity is, besides the safety of the therapy, the most important aspect of a drug product or therapy. Thus, a major goal of formulation development is to maintain the functionality of the therapeutic compound inside its formulation throughout its shelf-life. <br /><br />Functional titer, also known as infectious titer, is the measurement of how much virus actually infects a target cell. Functional titer may be expressed in the form of transduction units per mL (TU/mL), plaque-forming units per mL (pfu/mL), or infectious units per mL (ifu/mL), depending on the viral vector.




Principle

Quantitative reverse transcription PCR (RT-qPCR) is used when the starting material is RNA. In this method, RNA is first transcribed into complementary DNA (cDNA) by reverse transcriptase from total RNA or messenger RNA (mRNA). The cDNA is then used as the template for the qPCR reaction.

Applications

Biopharmaceutica

Procedure

1. Sample preparation.
2. Predenaturation (Optional) This step is recommended if the RNA template has a high degree of secondary structure.
3. Primer Extension. This step is recommended for extending primers.
4. cDNA Synthesis.
5. Reaction Termination.
6. Analyse the data.

Materials

• Sample: Peptides, Proteins, Vaccines, Virus-like particles
• Equipment: Quantitative Polymerase Chain Reaction (qPCR) instruments