In Vitro Biology

In Vitro Biology Services in India

Vipragen specializes in a broad range of laboratory assays, including biochemical, mechanistic, functional, and immunological assays. Our services cater to diverse research needs, offering tailored assays to meet unique client requirements. This comprehensive array ensures our team can provide valuable insights and solutions across various scientific domains.

Cell Biology

Vipragen's cell biology services deepen the understanding of how drug candidates interact with biological systems at cellular and molecular levels, crucial for target identification, mechanism elucidation, safety assessment, and successful drug discovery. Screening compounds in a multicellular humanized environment expedites data translation into clinical applications. We develop human primary cell-based assays, employing high-throughput methods across various disease indications, ensuring rapid identification of promising drug candidates within relevant biological environments.

Cell based & Cell free Assays

We've advanced from target-based screening in basic cell assays to using multicellular, disease-relevant platforms for functional and biochemical assays in drug discovery. Our tailored cell-based assays, crafted for every drug development phase, utilize diverse cell types and technologies to closely mimic specific disease states, enhancing translational success. Cell-based assays, conducted outside the living organism, measure cellular responses to stimuli, providing valuable insights into drug discovery, toxicity testing, and cellular functions. They offer a more complex and physiological context to assess the impact of substances on living cells.

Cell-Free Assays do not involve intact living cells; instead, they utilize extracted or synthesized cellular components like proteins, enzymes, or nucleic acids. These assays are valuable for studying specific biochemical reactions in a simplified and controlled environment, providing precision and convenience. They are commonly used in biochemical and drug discovery research to gain insights into the mechanisms of various biological reactions. In contrast, Cell-Based Assays involve intact living cells to study responses in a physiological context. Both assay types contribute to advancing our understanding of in vitro biology, playing crucial roles in drug development and fundamental biological research.

Molecular Biology Services in India

Vipragen provides customized and extensive molecular biology services, conducting a comprehensive range of genomics-based assays to support every stage of drug development, spanning from initial discovery and preclinical studies to clinical trials. Our services are adaptable to both single plex and multiplex formats, utilizing techniques such as qPCR and ddPCR. Additionally, our bioanalytical services cover the assessment of novel and intricate modalities, including siRNA, mRNA, and anti-sense oligos, employing viral vectors for nucleic acid therapeutics.

We specialize in isolating and purifying DNA and RNA from diverse sources such as tissues, body fluids, and cultured cells. Our extraction processes are meticulously optimized, incorporating automation to ensure the utmost quality. Prior to utilization, both the quality and yield of the extracted nucleic acid samples undergo thorough assessment, guaranteeing reliable and high-quality results.

Molecular Biology Services: A Diverse Range

Real-time Quantitative and Reverse Transcriptase PCR (qPCR, ddPCR):

Utilizing advanced techniques for precise quantification and analysis of nucleic acids in real-time and reverse transcription settings.

Nucleic Acid Therapeutics Assessment:

Conducting thorough evaluations, including biodistribution and shedding studies, along with quantifying transgene mRNA for gene and cell therapies.

Gene Expression Analysis:

Analyzing and interpreting gene expression patterns to understand the molecular mechanisms underlying various biological processes.

Cytokine mRNA Profiling in Immune Response:

Investigating and profiling cytokine mRNA expression to gain insights into immune responses and related pathways.

Biomarker Assessment:

Identifying and evaluating biomarkers that serve as indicators of biological processes, diseases, or drug responses.

CYP mRNA Induction:

Assessing the induction of cytochrome P450 mRNA, providing insights into drug metabolism and interactions.

Ex Vivo mRNA Profiling in Liver:

Profiling mRNA expression in liver tissue outside of the living organism to understand gene expression patterns.

Reporter Gene Assays:

Employing reporter genes to study the expression of specific genes and assess their response to various conditions.

Genomic and Plasmid DNA Isolation:

Isolating DNA from genomic and plasmid sources for downstream applications.

Residual Mammalian DNA Detection by Hybridization Analysis:

Detecting and quantifying residual mammalian DNA through hybridization techniques.

Total RNA Isolation:

Extracting RNA from diverse biological samples for downstream molecular analyses.

Assay Feasibility, Development, and Validation:

Assessing, creating, and validating assays to ensure accuracy and reliability in experimental outcomes.

Biological Sample Analysis:

Conducting in-depth analysis of biological samples to extract meaningful data and insights.

GLP/Non-GLP Study Designs and Reporting:

Designing and reporting studies adhering to Good Laboratory Practice (GLP) standards or non-GLP as required for regulatory compliance.

Microbiology

The Biologics Testing Solutions division at Vipragen delivers microbiology services to assist our clients in lot release programs, manufacturing, and stability testing requirements. Our testing adheres to various compendial methods specified by global regulatory bodies.

Sterile Products Testing Services

Sterile products, including medical devices, biologics, veterinary products, and pharmaceuticals labeled as sterile, undergo comprehensive testing to ensure quality and safety. Our testing services cover bioburden, sterility, and endotoxin assessments, adhering to global regulatory standards.

Testing Services:

Bioburden Testing (ISO 11737-1, USP <61>, USP <62>, EP 2.6.12, EP 2.6.13, JP 4.05):

Determine the quantity of viable aerobic microorganisms.

Establish the sterilization dose required for subsequent sterility testing.

Sterility Testing (EP 2.6.1, USP <71>, JP 4.06):

Compliance: Testing aligns with USP, EP, and FDA regulations.

Methods: Conducted through direct inoculation or membrane filtration, in isolator or cleanroom environments.

Bacteriostasis/Fungistasis Test: Assess whether the test article inhibits microbial growth.

Vaporized Hydrogen Peroxide (VHP) Ingress Testing: Evaluated for sterility tests within VHP-decontaminated isolators, ensuring the integrity of results.

Endotoxin Testing (EP 2.6.14, USP <85>, JP 4.01):

Methods: Gel-clot (qualitative), kinetic turbidimetric, and chromogenic (quantitative) assays.

Compliance: Meets pharmacopeia requirements.

Services: Preliminary screening, validation, and backup technical support provided.

Additional Services: In vivo pyrogen testing (EP 2.6.8, USP <151>) for materials unsuitable for or inadequately tested by endotoxin assays.

Monocyte activation testing (MAT) (EP 2.6.30) for cases where endotoxin testing is inappropriate or insufficient.

Our testing services ensure the sterility, safety, and compliance of sterile products, contributing to the overall quality assurance of pharmaceutical and medical devices.

Testing for Non-Sterile Products

Products that do not require sterility, including finished products and active pharmaceutical ingredients (APIs), undergo specific testing for bacterial and fungal counts, as well as the absence of pathogens. For certain products like creams and ointments, additional preservative efficacy testing may be necessary.

Testing Procedures:

Microbial Limits Test (EP 2.6.12, EP 2.6.13, USP <61>, USP <62>, JP 4.05):

Quantitatively enumerate bacteria and fungi that may grow under aerobic conditions.

Assess the microbial content of non-sterile products, ensuring compliance with specified standards.

Preservative Efficacy Testing (EP 5.1.3, USP <51>):

Antimicrobial preservatives are added to pharmaceutical preparations lacking sufficient antimicrobial activity.

Demonstrate the efficacy of the antimicrobial preservative through testing.

Ensures that the added preservative effectively prevents microbial growth, maintaining product integrity.

These testing procedures for non-sterile products are essential for confirming the microbiological quality and safety of finished pharmaceuticals and active ingredients. Additionally, preservative efficacy testing is crucial for products requiring antimicrobial protection, such as creams and ointments.

Additional Microbiology Testing Services

Our microbiology testing services extend beyond sterility assessments for various products, encompassing a diverse range of specialized analyses:

Cytotoxicity Testing (ISO10993 Part 5, USP <87>):

Detect the presence of toxins in medical devices, their components, or raw materials.

Water Analysis:

Provide total viable and coliform organism counts necessary for assessing process water quality.

Mycoplasma Testing:

Determine the presence of mycoplasma in a test article using agar-cultivable, non-cultivable, and PCR methods.

Mycoplasmastasis:

Assess if a test article inhibits the growth of mycoplasma, aligning with EU and USP requirements.

Spiroplasma Testing (PCR):

Detect the presence of spiroplasma in a test article, particularly applicable to products produced in insect and plant cells.

Mycobacterium Testing:

Determine the presence of mycobacterium species in the test article, crucial for cell-derived vaccine products.

Disinfectant Testing (Biocidal Products Directive - BPD-98/8/EC):

Verify the efficacy of disinfectant programs for cleanrooms or controlled areas.

Antibiotic Testing (EP and USP Specifications):

Evaluate the potency or activity of an antibiotic by assessing its inhibitory effect on microorganisms.

Environmental Monitoring:

Offered to companies with on-site cleanrooms of various grades.

Vipragen can conduct monitoring or train operational staff, including sample incubation and reporting.

These specialized services contribute to the overall quality control and safety assessment of diverse products and environments in the pharmaceutical and medical industries.

Microbial Exploration Initiatives

The microbiology group's second critical role involves furnishing bacterial challenge material for vaccine studies. This encompasses exploratory work on bacterial isolates, the meticulous preparation of challenge materials, and the subsequent analysis of organs and blood samples upon the study's conclusion. All these activities are conducted within specialized and dedicated facilities

Biopotency and Bioassays

Bioactivity & Potency Assays

Potency testing is a crucial aspect ensuring the quality, safety, and efficacy of biopharmaceutical products, along with confirming biocomparability between innovator and biosimilar products. Our comprehensive array of potency testing services for various biologics encompasses both in vivo and in vitro bioassays.

Potency Testing and Bioassay Development Services:

Method Development, Transfer, and Optimization:

Develop, transfer, and optimize methods for effective potency testing.

Tailoring methodologies to specific requirements, ensuring accuracy and reliability.

Phase-Appropriate Method Validation:

Validate methods at relevant phases of development.

Ensuring method reliability aligns with the current stage of product development.

Lot Release Testing for Drug Substance and Drug Product:

Ensuring consistency and adherence to defined standards for both drug substance and drug product.

Assess the quality and potency of each production lot before release.

Stability Storage and Testing:

Evaluate product stability under defined storage conditions.

Ensuring product efficacy and quality over its designated shelf life.

Accelerated Stress Studies:

Subject products to accelerated stress conditions.

Assessing product stability and potential issues under extreme conditions.

Comparability Testing Between Innovator and Biosimilar Biological Products:

Conduct testing to compare innovator and biosimilar products.

Ensuring bioequivalence and similarity between the reference and biosimilar products.

These services encompass the development, validation, and application of methods for potency testing, ensuring the quality, safety, and efficacy of biopharmaceutical products throughout their lifecycle.

In Vitro Bioassays

In vitro bioassays are essential for maintaining the quality, safety, and effectiveness of biopharmaceutical products. They play a critical role in verifying bio comparability between innovator and biosimilar products. Ensuring the reliability, standardization, and relevance of these assays is crucial for an accurate representation of the product's mode of action. Bioassays are central to potency determination for regulatory submission and lot release, making them critical for product development and manufacturing. They are necessary to confirm the continued quality, safety, and efficacy of biopharmaceutical products, as well as to establish bio comparability between innovator and biosimilar products. The selected assays must be reliable, standardized, and relevant to accurately reflect the product’s mode of action.

Product-Specific Bioassays for Antiviral Compounds (Interferons)

Bioassays specific to antiviral compounds, particularly interferons (IFN-α, IFN-β), have been conducted for over a decade. These compendial bioassays are grounded in assessing the inhibitory activity of interferons against the cytopathic effect induced by a virus on a susceptible cell line. All bioassays are in full compliance with the stipulations outlined in the European Pharmacopoeia and have undergone validation in accordance with ICHQ2(R1) guidelines.

Potency Assessment of Growth Factors

The effectiveness of human growth factors, including EPO, GM-CSF, and G-CSF, is evaluated through conventional proliferation assays. These assays have demonstrated success in their application to both originator and first- and second-generation biosimilar products. When applicable, the assays adhere to the standards set by the European Pharmacopoeia, and each assay has undergone validation in accordance with the guidelines outlined in ICHQ2(R1).

Hormones: Parathyroid Hormone (PTH) Assessment

A cell-based assay is conducted for parathyroid hormone (PTH), utilizing the determination of cyclic AMP (cAMP) release. This process is detected through homogeneous time-resolved fluorescence (HTRF) or ELISA. The methodology employed has undergone validation in accordance with the guidelines outlined in ICHQ2(R1).

Monoclonal Antibodies: Antibody-Dependent Cell Cytotoxicity (ADCC) Assessment

Antibody-dependent cell cytotoxicity (ADCC) is evaluated by measuring LDH release. This is accomplished using NK (Natural Killer) effector cells that are freshly isolated from peripheral blood mononuclear cells (PBMCs) or through a reporter-based bioassay. The selection of the target cell line is tailored to the specific product under examination.

Antibody-Dependent Cellular Phagocytosis (ADCP) Assessment

The measurement of antibody-dependent cellular phagocytosis (ADCP) is conducted using either a luminescence-based reporter bioassay or flow cytometry. Macrophages for this analysis are differentiated from monocytes isolated from peripheral blood mononuclear cells (PBMCs).

Complement-Dependent Cytotoxicity (CDC) Assessment

The evaluation of complement-dependent cytotoxicity (CDC) is performed utilizing flow cytometry, employing a live/dead discrimination dye or a luminescence-based approach. In this process, the antibody marks an appropriate target cell line, which is subsequently targeted and attacked by the complement cascade.

Apoptosis/Programmed Cell Death (PCD) Assessment

The investigation of apoptosis or programmed cell death (PCD) is commonly conducted through either a reporter gene assay or flow cytometry-based assays.

Mode of Action Assays for Monoclonal Antibody Therapeutics:

Assessing how monoclonal antibody therapeutics function.

Antigen, Receptor, or Ligand Density (e.g., Binding Assays and Competitive Binding Assays):

Evaluating the density of antigens, receptors, or ligands through binding assays.

Multiplexing Analyses of Cytokines (CBA Technology):

Utilizing CBA technology for simultaneous analysis of multiple cytokines.

Cell-Based Immunogenicity:

Evaluating immunogenicity through cell-based approaches.

Intracellular Protein Expression:

Analyzing the expression of proteins within cells.

Transgenic Products In Vivo (e.g., Green Fluorescent Protein - GFP):

Assessing products, such as GFP, expressed in living organisms.

Enzyme Activity:

Measuring the activity of enzymes.

Phosphoprotein Analysis:

Analyzing proteins based on phosphorylation.

Apoptosis/Viability:

Assessing programmed cell death and overall cell viability.

Cell Cycle Analyses:

Studying changes in the cell cycle.

Changes in Intracellular pH, Calcium, and Glutathione:

Monitoring variations in intracellular pH, calcium levels, and glutathione concentrations.

Various Combinations (DNA/Surface Antigens, etc.):

Examining diverse combinations, such as DNA and surface antigens.

In-Process Quality Control of Primary Cells:

Performing quality control checks during the production process of primary cells.

Time-Resolved Fluorescence:

The time-resolved fluorescence method relies on fluorescence resonance energy transfer (FRET) within a microtiter plate. Frequently employed for third-generation anticancer and anti-inflammatory drugs, this method is effective in elucidating the activation of specific phosphorylation pathways in target cells. To validate the mode of action of these drugs, the assay must accurately depict the impact on the phosphorylation of key mediators within the involved pathway.

Advantages of the Method:

Low Background: Minimal interference from background signals.

Increased Assay Sensitivity: Enhanced sensitivity compared to conventional phosphorylation determination methods like ELISA.

Reduced False-Positive or False-Negative Results: Fewer instances of inaccurate positive or negative findings.

Suitability for Cell-Based Assays: Applicability to assays conducted with cells.

Homogeneous Time-Resolved Fluorescence (HTRF) Technology:

Homogeneous time-resolved fluorescence (HTRF) technology presents an intriguing alternative for mode of action assays, offering advantages such as low background, heightened sensitivity, and compatibility with cell-based assays.

Target-Specific Reporter Bioassays:

For monoclonal antibody therapeutics deviating from classical mechanism of action (MOA) pathways, target-specific reporter solutions are available. An example includes solutions tailored for anti-VEGF antibody therapeutics.

Biomarkers

Biomarker assays are crucial not only for evaluating therapeutic candidate activity but also for target validation, supporting mechanisms of action, predicting therapeutic efficacy, and early toxicity identification. Discovering and validating disease-related and pharmacodynamic biomarkers is a complex process that starts early in drug discovery, shaping a robust testing strategy across development phases. Our global biomarker services cover safety assessments in diverse therapeutic areas, translating biomarkers seamlessly into clinical applications, expediting candidate development, and reducing timelines. Leveraging genomics, transcriptomics, proteomics, flow cytometry, and metabolomics, our approach enables diverse biomarker discovery and development avenues.

Biomarker Discovery Services:

Our proficiency and knowledge in biomarker discovery services empower us to contribute to the identification of biologically and scientifically relevant biomarkers. By measuring and interpreting changes within the test system and correlating them with the drug development program, we ensure scientific excellence. This is achieved through a collaborative synergy among experts in various domains, adding substantial value to the studies. From assay qualification to comprehensive validation, our experts propose fit-for-purpose validation methods tailored to the context of use (COU) of your biomarkers.

Identification of Predictive Biomarkers:

The discovery and utilization of predictive biomarker assays have become integral components of the drug discovery and development process. Vipragen boasts extensive expertise in biomarker identification and assay development across diverse therapeutic areas, including respiratory disease, oncology, CNS, and metabolic diseases.

Biomarker Analysis Services:

Biomarker assays and analytical readouts serve as vital tools in drug discovery, spanning target validation studies to candidate selection. The ability to measure with precision and sensitivity is crucial for making critical go/no-go study decisions. With years of experience in assay design and sample processing, Vipragen provides support to your drug discovery pipeline by delivering biomarker analysis services in a non-GLP environment. Whether your study is conducted at your facility or designed and implemented at ours, our focus on strategic assay validation/qualification ensures accurate testing of study samples, offering reliable biological translation to your drug discovery program.

In-vitro Efficacy Models

Nutraceuticals are bioactive compounds found in foods or supplements that can have potential health benefits beyond their basic nutritional value. When studying the efficacy of nutraceuticals for specific health conditions like obesity, cancer, diabetes, liver toxicity, and cognition, researchers often use both in-vivo and in-vitro models to assess their effects. Let's look at each of these areas in more detail:

Anti-Obesity:

In-vitro studies use cells or tissues grown in a controlled environment. Researchers can use adipocyte cell lines to investigate the direct impact of nutraceuticals on fat storage, lipolysis, and adipokine production.

Anti-Cancer:

In-vitro models use cancer cell lines to evaluate nutraceuticals' effects on cell proliferation, apoptosis, and the inhibition of cancer pathways.

Anti-Diabetic:

In-vitro studies can use insulin-secreting cell lines (e.g., pancreatic beta cells) to assess the impact of nutraceuticals on insulin secretion and glucose utilization.

Liver Toxicity:

Liver cell cultures or hepatocyte cell lines can be used to assess the impact of nutraceuticals on liver enzyme activity, detoxification processes, and oxidative stress.

Cognition:

In-vitro neuronal cell cultures assess nutraceutical impact on neuronal growth, synaptic plasticity, and neuroprotection. Both in-vivo and in-vitro models have advantages and limitations. In-vivo models mimic living organisms but can be costly and ethically challenging, while in-vitro models offer controlled environments for detailed studies but lack full human body complexity. Model choice depends on the research question and ethics, with researchers often combining in-vivo and in-vitro models for a comprehensive understanding of nutraceutical efficacy.

Claim Substantiation and Efficacy Testing:

Claim substantiation and efficacy testing of Vipragen are essential processes in the development, marketing, and regulation of nutraceuticals, which are products that combine the characteristics of both nutrition and pharmaceuticals. These processes are crucial for ensuring that the claims made about nutraceutical products are accurate and supported by scientific evidence. Here's an explanation of these terms and their significance:

Claim Substantiation:

Claim substantiation involves providing scientific evidence to support nutraceutical product claims, covering health benefits, nutritional content, or other promoted attributes. Nutraceutical companies must furnish credible data, such as clinical trials and research studies, to validate their claims. Vipragen emphasizes accurate and truthful consumer information, aiming to prevent misleading content. This process is essential for regulatory compliance and building consumer trust.

Efficacy Testing:

Efficacy testing assesses a nutraceutical's ability to deliver claimed health benefits or address specific concerns. Typically involving clinical trials or laboratory experiments, these studies aim to demonstrate the product's impact on targeted health outcomes. Such testing is crucial for regulatory approval and marketing, confirming the product's effectiveness in fulfilling its intended purpose.