Biosimilar Testing Services

Reach us to better understanding!!

Biosimilar testing services

What are Biosimilars?

A biosimilar is a biological product that is highly similar to and has no clinically meaningful differences from an existing FDA-approved reference product. Biosimilars are known as similar biologics, follow-up biologic, similar biological products, subsequent entry biological, second entry biological, biogenerics, multisource products, similar biotherapeutic product and off-patent biotech products. The nomenclature varies from one geography to another. 

A reference product is the single biological product, already approved by FDA, against which a proposed biosimilar product is compared. A reference product is approved based on, among other things, a full complement of safety and effectiveness data.  Reference product is also known as Innovator product, Originator drug, Novel Biologic and Reference Medicinal Product.

The first biosimilar approved by EMA was Omnitrope in 2005; US FDA approved Zarxio its first biosimilar in 2015. The first ‘similar biologic’ to be approved and marketed in India was for  hepatitis B vaccine in 2000, in recent years almost 98 similar biologics are approved or are under development in India.

Difference between Novel Biologics and Biosimilars

Novel Biologics 

Biosimilars

Novel therapeutic

Copy of Novel Biologics

Time to market- more

Time to market- comparatively lesser

Patentable

Non-patentable

Price is higher

Price is comparatively lower

Extensive clinical studies required

Comparatively lesser clinical studies required

Examples of few Biosimilars

Novel Biologics 

Generic name

Biosimilars

Humira (AbbVie)

Adalimumab

ZRC3197 (Adalimumab Biosimilar, Cadila), Cyltezo (Boehringer Ingelheim)

Rituxan (Genentech)

Rituximab 

Rixathon® [Sandoz rituximab], Truxima® [Celltrion rituximab]

Herceptin (Genentech)

Trasuzumab 

CanMab/Hertraz(Biocon/Mylan, India*/USA, Hervycta (DRL), 

Neupogen (Amgen)

Filgrastim

Zarxio (Sandoz), Nufil (Biocon)

Remicade (Janssen Biotech)

Infliximab

Inflectra (infliximab-dyyb, Celltrion) and Renflexis (infliximab-abda, Samsung Bioepis)

What is BPCI Act?

The aim of the Biologics Price Competition and Innovation (BPCI) Act BPCI Act is similar, in concept, to that of the Drug Price Competition and Patent Term Restoration Act of 1984 (a.k.a the “Hatch-Waxman Act”) which created abbreviated pathways for the approval of drug products.  Under the BPCI Act, a sponsor may seek approval of a “biosimilar” product under new section 351(k) of the PHS Act.  A biological product may be demonstrated to be “biosimilar” if data show that the product is “highly similar” to the reference product notwithstanding minor differences in clinically inactive components and there are no clinically meaningful differences between the biological product and the reference product in terms of safety, purity and potency.

Biologics Price Competition and Innovation (BPCI) Act

  • Created provision for abbreviated approval pathway for biological products; 351(k)
  • To provide the public with greater access to safe and effective biological products.
  • This allows for a potentially shorter and less costly drug development program for a biosimilar.

What is Biosimilar Development Program?

The goal of a biosimilar development program is to demonstrate biosimilarity between the proposed biosimilar product and the reference product, not to independently establish the safety and effectiveness of the proposed product. This generally means that biosimilar manufacturers do not need to conduct as many expensive and lengthy clinical trials, potentially leading to faster access to these products, additional therapeutic options, and reduced costs for patients.

A biosimilar product application must include data demonstrating biosimilarity to the reference product. This usually includes data from:

  1. Analytical studies demonstrating that the biological product is highly similar to the reference product, notwithstanding minor differences in clinically inactive components;
  2. Animal studies, including an assessment of toxicity; and
  3. A clinical study or studies sufficient to demonstrate safety, purity, and potency of the proposed biosimilar product in one or more of the indications for which the reference product is licensed. This typically includes assessing immunogenicity, pharmacokinetics (PK), and, in some cases, pharmacodynamics (PD) and may also include a comparative clinical study. 

The guidelines regulating the Biosimilar development are EMEA in Europe, US-FDA in US, CDSCO-RCGM in India and WHO. Extensive analytical characterization is required to establish biosimilarity. The following table describes the different tests which are required for biosimilairty demonstration. Currently, CBA is well equipped to provide almost 80 percent of the testing services (as highlighted in green boxes) required for biosimilarity assessment. Kindly note the animal or in-vivo studies are out of scope of CBA.

CBA offers Biosimilar product characterization, Innovator product characterization, Batch release assays, and stability studies as a part of the Biosimilarity testing service offering. To know more about CBA capabilities and services, please visit: https://bioanalysis.in/protein-characterization-services/

CDSCO

EMEA

USFDA

ICH Q6B

Amino acid sequence (full as well as N and / or C terminal) 

Amino acid sequence (full as well as N and / or C terminal) 

Primary structure analysis- Amino acid sequence 

Amino acid sequence

Intact Mass assessment by LC-ESI-MS

Intact Mass assessment by LC-ESI-MS

Higher order structure analysis- Secondary, Tertiary, Quaternary (including aggregation)

Amino acid composition

Peptide mapping

Peptide mapping

Enzymatic posttranslational modifications, such as glycosylation and phosphorylation

Terminal amino acid sequence

Far UV Circular Dichroism Spectrum

Sulfydryl groups(s) and disulphide bridges

Other potential variations, such as protein deamidation and oxidation

Peptide map

FTIR Analysis

N-terminal sequence confirmation

Intentional chemical modifications, such as PEGylation sites and characteristics

Sulfhydryl group(s) and disulfide bridges

Tryptic Peptide Map-1D and 2D

C-Terminal sequence confirmation

Effect of excipients,
Purity,
Process impurities
Product impurities
Stability

Molecular weight or size

Sulfydryl groups(s) and disulphide bridges

Glycan analysis- degree of mannosylation, galactosylation, fucosylation and sialylation. Distribution of the main glycan structures present (often G0, G1 and G2) should be determined

Receptor Binding assay

Isoform pattern

Fluorescence spectrum

Carbohydrate content

In-vitro bioassay

Extinction coefficient (or molar absorptivity)

Near UV Circular Dichroism

Higher order structure (using appropriate techniques)

Apoptosis assay (if applicable)

Electrophoretic patterns

UV-VIS spectroscopy

Fluorescence spectrum

Neonatal Receptor(FcRn) Binding Assay

Liquid chromatographic patterns

Glycoforms and other modificaions like phosohorylation, acetylation,
myrsitoylation,esterification by HPLC& MALDI-TOF

Circular Dichroism

ADCC

Spectroscopic profiles

Isoforms and charge variants by Isoelectricfocusing

Receptor Binding assay

CDC

Cell substrate-derived impurities

N-terminal sequence confirmation

CDR identification

Neutralizing Biological activity in actual target host cell

Cell culture-derived impurities

C-Terminal sequence confirmation

Receptor Binding assay (FcRs)

Kinetics of neutralizing biological activity during the proposed therapeutic period of protection

Downstream-derived impurities

Ion exchange chromatography for charge heterogeneity

ADCC

Efficacy in appropriate disease/ infection model in-vitro and/or in-vivo

Truncated forms

Product related variants and impurities by RPLC, SE-LC (CQA), IE-LC, SDS-PAGE, IEF, Western Blot (CQA)

CDC

In-vivo bioassay(if available)

Other modified forms

Host cell protein analysis
• Host cell DNA analysis
• Residual ProteinA (if applicable)
• Pyrogen content

Apoptosis assay (if applicable)

 

Aggregates

Protein content(CQA)
Appearance(KQA)
pH(KQA)
Osmolarity(KQA)
Composition of key excipients including stabilizer(if formulation is same) (KQA)
Visible/subvisible particles,(KQA)
Pyrogen content(KQA)

Neonatal Receptor(FcRn) Binding Assay

 

Animal-based biological assays

Receptor Binding assay

In-vivo bioassay(if available)(KQA)

 

Cell culture-based biological assays

In-vitro bioassay

Molecular weight or size, isoform pattern, extinction coefficient, electrophoretic profiles, chromatographic data and spectroscopic profiles.

 

Biochemical assays

Apoptosis assay (if applicable)

HCP

 

Purity

Neonatal Receptor(FcRn) Binding Assay

HCD

 

Impurities

Receptor Binding assay (FcRs)

Cell culture residues, downstream processing residues

 

Contaminants

ADCC

Contaminants- microbial species, endotoxins

 

Neutralizing Biological activity in closest animal species

CDC

appropriate physicochemical and/or immunochemical assay

 

Kinetics of neutralizing biological activity during the proposed therapeutic period of protection

Neutralizing Biological activity in actual target host cell

  

Efficacy in appropriate disease/ infection model in-vitro and/or in-vivo

Neutralizing Biological activity in closest animal species

  

In-vivo bioassay(if available)(KQA)

Kinetics of neutralizing biological activity during the proposed therapeutic period of protection

   

Efficacy in appropriate disease/ infection model in-vitro and/or in-vivo

   

In-vivo bioassay(if available)(KQA)

   

References: