Gene Expression Analysis by qPCR (RT-qPCR)
We develop and perform RT-qPCR assays for the quantification of gene expression, biomarkers, and transgenes, generating sensitive and reproducible data for preclinical, clinical, and bioanalytical studies.
- Gene expression and biomarkers
- Transgene and viral vector quantification
- Method development and validation
- Regulatory support
We offer molecular biology solutions tailored to each product, platform, and study objective, enabling the quantification of relevant gene signals in different biological matrices using robust and highly sensitive strategies.
We work from method design and control selection through to data analysis and interpretation, providing useful tools for studies of biological response, advanced therapies, and analytical control.
We apply these capabilities to the quantification of biomarkers, transgenes, and viral vectors, with methods adapted to each product and analytical support useful for development, characterization, and control in complex biological platforms.
What is gene expression analysis by qPCR?
qPCR (quantitative real-time PCR) is a technique based on the enzymatic amplification of nucleic acids and their real-time detection, allowing precise quantification of RNA after its conversion into cDNA.
This approach enables the evaluation of changes in gene expression associated with:
- Drug response
- Inflammatory and immune processes
- Biomarkers of efficacy or toxicity
The challenge in gene expression studies
If these factors are not properly controlled, they may compromise the biological interpretation of results. Gene expression measurement presents critical technical challenges:
- RNA degradation
- Variability in reverse transcription
- Differences between biological matrices
- Amplification efficiency
Our solution: High-precision RT-qPCR
At Netpharmalab, we implement RT-qPCR assays in the following formats:
- SYBR Green
- Specific probes (TaqMan®)
Ensuring maximum quality through:
- Standardized RNA extraction
- Use of reference genes (housekeeping genes)
- Validation of amplification efficiency
- Normalization using ΔCt, ΔΔCt, or absolute quantification
Main applications
- Validation of response and toxicity biomarkers
- Immune and inflammatory response studies
- Transgene and viral vector quantification
- Evaluation of drug-induced expression
- Stability and biological potency studies
- Support for PK/PD studies and correlation with ADA/NAb
What does our service include?
- Messenger RNA (mRNA) quantification
- Gene biomarker analysis
- Expression studies in multiple matrices
- Customized methods according to the study
- Quantitative and reproducible results
RT-qPCR Methodology
- RNA extraction
From plasma, serum, tissues, cell cultures, or biological products - cDNA synthesis
Reverse transcription using random primers or oligo-dT - Real-time amplification
Using SYBR Green or TaqMan® probes - Quantification
– Standard curves
– ΔCt / ΔΔCt
– Absolute quantification - Quality controls
– No-RT
– No-template
– Housekeeping genes (GAPDH, ACTB, etc.)
Assay phases
- Method design / transfer
- Qualification and analytical validation
- Sample processing
- Data analysis and reporting
Including evaluation of:
- Linearity
- Precision
- Reproducibility
- Limit of detection
Regulatory compliance
With complete SOPs and traceability at all stages of the process, our assays comply with:
- EMA (European Medicines Agency)
- FDA (Food and Drug Administration)
- ICH Q2(R2)
- 21 CFR Part 11 (data integrity)
Frequently Asked Questions (FAQ)
- What types of samples can be analyzed?: Plasma, serum, tissues, cell cultures, and biological products.
- What is the difference between SYBR Green and TaqMan®?: SYBR Green detects DNA in a general manner, while TaqMan® provides greater specificity through probes.
- What is ΔCt or ΔΔCt normalization?: These are methods used to compare gene expression levels by adjusting results against reference genes.
- Can transgenes or viral vectors be quantified?: Yes, qPCR is a key tool for the precise quantification of genetic material.
- Are these assays valid for regulatory studies?: Yes, provided they are developed and validated according to EMA, FDA, and ICH guidelines.
Request more information about our biological product analysis services
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