Biopharmaceuticals

Biopharmaceuticals

Bioneeds is dedicated to provide end-to-end Biologics Drug Development Services with its two distinct technology platforms:



Capabilities include development of recombinant proteins such as non-glycosylated proteins; glycoproteins and monoclonal antibodies derived from either bacterial or mammalian host expression systems.

Our Preclinical platform is supported by OECD-GLP and AAALAC certified animal facilities.

We have the expertise to handle Novel Biologics, Biosimilars and Biobetters.

CQA Driven Process & Product Development

  • Cell Line Development

  • Cell Banking & Characterization

  • Upstream Process Development

  • Downstream Process Development

  • Formulation & Stability

Well Characterized Products

  • Structure

  • Function (MOA)

  • Safety

  • Toxicology

  • Immunogenicity

Preclinical Studies:

  • Animal Toxicity

  • PK/PD

  • Immunogenicity

  • POC studies MOA


Preclinical Platform



Pre-clinical Toxicology (GLP/Non GLP)

RCGM; EMEA; US-FDA

Toxicological Studies


Acute Toxicity:

  • Route of Administration: IM/IV/SC

  • Animal models: Rats/Rabbits/Mice

Repeated Dose Toxicity:

  • Route of Administration: IM/IV/SC

  • Animal Species: Rats/Rabbits

Dermal Irritation Test:

  • Procedure: Dermal Patch

  • Animal Species: New Zealand White Rabbits

Skin Sensitization Test:

  • Procedure: Guinea Pig Maximization Test (GPMT)

  • Animal Species: Guinea Pig

Guidelines:

  • EMEA-CHMP
  • CDSCO
  • US-FDA
  • ICH

IBSC Approved Protocols GLP/Non GLP Studies NOAEL Clinical Chemistry Biochemical Analyses Histopathology Animal Health

Note: Other route of administration routinely used: Intra-ocular/Intra-peritoneal/Intra-dermal


Bioassays (in vitro/in vivo): (GLP/Non GLP)

Method Development; Validation;
Sample Analysis

Bioassays

Proof-of-Concept (POC); Mechanism of Action (MOA); Potency


In vitro Assays:

Cell Based Bioassays

  • Proliferation assays

  • Cytopathic Effect Reduction Assays (CPER) for Interferon

  • CDC assay

  • ADCC assay

  • Cytokine Release Assays

  • Reporter Gene Assays

Binding Kinetics

  • ELISA based assays

  • BIACore based assays

  • Flow Cytometry based assays

In vivo Assays:

Retic count Assay for Erythropoietin; Darbapoetin and other NESP Ovary/Seminal Vesicle weight gain assay for FSH; HCG and LH Absolute Neutrophil Count Assays for GCSF/Peg GCSF Glucose Uptake assay for Insulin

Guidelines:

  • USP/EP/IP
  • Monographs
  • USP<111>
  • USP<1033>
  • ICH

Parallel Line Analysis 4PL Analysis Signal/Noise Precision/Accuracy Linearity Specificity Confidence Limits


PK/TK Analysis (GLP/Non GLP)

Animal Studies; Sample Analysis

PK/TK Studies


Pharmacokinetics (PK) Studies:

  • Animal Species: Rat/Rabbits/Mice

  • Drug Dosing and Sampling: Based on design

  • Analyte Estimation: ELISA

  • Data Analysis: WinNonLin/SPSS

Toxicokinetics (TK) Studies:

The application of PK principles in animal toxicity studies provides information on exposure, half-life and elimination of the parent compound.

These studies are done in conjunction with repeated dose toxicology studies

Guidelines:

  • CDSCO
  • US-FDA
  • ICH

Bioequivalence PK Parameters Cmax ; t1/2 ; AUC etc. Signal/Noise MRD Precision/Accuracy Linearity Specificity Stability Confidence Limits



Immunogenicity Testing (GLP/Non GLP)

Antibody Generation; Screening Assays; Confirmatory Assays;
Neutralizing Antibody Assays

Immunogenicity Testing (GLP/Non GLP)

Antibody Generation; Screening Assays; Confirmatory Assays;
Neutralizing Antibody Assays


Animal study: Drug Dosing & Sampling

  • Animal Species: Rat/Rabbits/Mice

Drug Specific Antibody Generation

  • Animal Species: Rat/Rabbits/Goat/Mice

  • Purification and Characterization

Assays for Immune Response

  • Screening Assay (ELISA based)

  • Confirmatory Assay (ELISA based)

  • Neutralizing Assay (Cell Based)

Immunogenicity Testing


Impurity Analysis (GLP/Non GLP)

HCP; HCD; Others

Impurity Analysis

Host Cell DNA (HCD) & Host Cell Protein (HCP)


  • Host Cell DNA (Microbial & Mammalian) qPCR

  • Host Cell Protein (Microbial & Mammalian) ELISA

  • Process Specific HCP Assay Development Cascading Immunization

Guidelines:

  • CDER/CBER
  • USP<1132>
  • USP<1010>
  • ICH

Signal/Noise Slope/PCR efficiency Precision/Accuracy Linearity Specificity Confidence Limits


Antibody Development (Monoclonal/Polyclonal)

Immunization; Purification; Characterization; Reactivity

Antibody Development


Polyclonal Antibody Generation

  • Animal Species: Rat/Rabbits/Goat

Monoclonal Antibody Generation

  • Animal Species: Rat; Mice

Antibody fragments

  • by Enzyme treatment

Purification and Characterization

  • Protein A/Protein G (Affinity Chromatography)

  • Purity: >90% (SDS-PAGE; 2DGE)

  • Quality: Very high specific reactivity (ELISA; Western Blot)

Experience in Antibody Generation:

  • GCSF
  • Peg-GCSF
  • FSH
  • Etanercept
  • HCG
  • Insulin
  • Exenatide
  • Asparaginase
  • Peg-Asparaginase


Drug Development Platform



Cell Line Development

Codon Optimization; Clone Stability; Bioreactor Suitability; Titre ; Product Quality

Cell Line Development


1. Host

  • Suspension CHO

  • cGMP grade

  • CD media

  • Non-animal origin media

  • Serum free

  • Complete traceability

2. Gene Synthesis

  • Sequence identification

  • Codon Optimization

  • Gene synthesis

  • Expression construct

  • Sequence confirmation

3. Transient Transfection

  • Transient transfection

  • Pools generation

  • Selection & amplification

  • Productivity assessment

  • Leader sequence processing

  • CQA evaluation of pools for shortlisting

4. Single Cell Cloning

  • Extensive screening

  • Monoclonality documentation

  • Growth kinetics

  • Specific productivity

  • Product quality & titer evaluation

5. Primary Cell Bank

  • Bacterial, fungal & mycoplasma testing

  • Gene copy numbers

  • Scale-up/ Bioreactor suitability

  • Stability till 60 generations

  • Cell line development documentation

6. MCB & WCB

  • Protocol evaluation

  • System suitability checking

  • Results verification

  • US/EU/JP guidelines


Process Development

Cell Culture & Purification ; Critical Process Parameters; COGS

Process Development


Process Developement


Product Characterization

Analytical; Bio/Analytical; Potency (Compliance to Standard Guidelines and Pharmacopeia)

Analytical & Structural

Primary Structure


AA Sequence & Variants

Intact Mass:

  • ESI-MS

  • Glycan removal by PNGaseF

Bottom-up Peptide Mapping:

  • Enzyme digestion and LC-MS/MS

Middle-up LC-MS:

  • Reduced Mab (LC-HC)

  • Limited proteolysis

Top-Middle-Down:

  • HR-MS/MS

SEC

CE-SDS

Charge Variants

Separation:

  • IEF, cIEF

  • HPLC (IEX, RP, HIC)

Mass Spectrometry:

  • Peptide Mapping (LC-MS/MS)

  • Top-down MS/MS

  • Middle up LC-MS

Glycovariants

Glycan:

Released:

  • CE-LIF

  • HPLC (NP- HILIC, IEX)

  • MALDI-TOF, ESI-MS, MS/MS

  • Intact middle-up LC-MS

  • Peptide mapping (LC-MS/MS)

Cysteine Linked Variants

Ellman Assay for free Cysteine:

Peptide Mapping:

  • Reduced & Non reduced

  • CID & ETD (Cys linkage)

Higher Order Structures, Aggregates & Complexes


Higher Order

  • Native-MS

  • Ion-mobility-MS (IM-MS)

  • HDX-MS

  • CD

  • Fluorescence

Aggregates

  • SEC

  • AUC

  • Native-MS

  • IM-MS

  • HDX-MS

  • DLC

Complexes

  • SPR

  • ELISA

  • FACS

Stability


Purified Protein PBS/Buffers

Thermal Stability DS/DP*

Real Time Real Temperature

Accelerated Stability

Stressed Stability

Freeze/Thaw

* DS: Drug Substance

* DP: Drug Product

Photo Stability

Serum Stability

Protein Concentration; Charge Variants; Aggregates & Degradation; Potency

HPLC (SEC;RP); CE-SDS; SDS-PAGE ; CEX; HIC; in Vitro/in Vivo Bioassay



Product Release Testing

Compliance to Standard Guidelines and Pharmacopeia

Product Release Testing


pH

  • Physical appearance

  • Osmolality

Identity

  • Immunoblotting

  • Isoelectric focusing

  • Bioassay

  • SEC

  • Peptide mapping

Assay

  • Cell based bioassay

  • ELISA based binding assay

  • SPR based assay

Concentration

  • RP-HPLC

  • SE-HPLC

  • OD

Purity

  • Ion exchange

  • RP-HPLC

  • SE-HPLC

  • HIC

Safety

  • Sterility

  • BET

  • Bioburden


Sequence to Clinic (Pre-IND Package)

Complete Product Development (starting with sequence) with tox material supply

Pre-IND Package

CQA Based Process & Product Development



  • Gene Sequence

  • Cell Line

  • Upstream Process

  • Down- stream Process

  • Formulation Stability Studies

  • Pre-Clinical Studies



  • Sequence verification Cell Line Stability Product CQA Productivity

  • Critical Process Parameters CQA/Characterization/Specification Stability COGs

  • Regulatory Compliance - Preclinical Studies Pre-IND filling


Our Flexible business models help minimize your development risk and conserve your R&D budget, which leads to successful development outcomes




Contact Us @

Phone  +91 80 22658400
+91 816 2214400
+91 98444 57677

Mail Us @

Email  bioneeds@bioneeds.in

Enquiry Request