The global pharmaceutical analytical testing outsourcing market was estimated at $6.1 billion in 2019 and is anticipated to register a CAGR of 8.3% through 2027, according to Grand View Research, Inc.¹ Increasing demand for analytical drugs, biosimilars, and biopharmaceuticals are contributing to market growth. 

Other factors such as increasing investments in R&D for pharmaceuticals, rising demand for product safety and quality, and changing regulations for in vivo and in vitro tests are also expected to drive the demand for pharmaceutical analytical testing outsourcing services. Additionally, the development of combination products, biosimilars, and other innovative medicines has led to an increase in demand for specific types of pharmaceutical analytical tests, such as bioana­lytical testing, method development and validation, active pharmaceutical ingredient testing, and stability testing.

Demonstrate Control Over Biologics Manufacturing

Analytical testing of biologics presents unique challenges relative to small-molecule APIs and drug prod­ucts. Manufacturing biologics using cellular systems results in a mixture of molecules from the expression of the biologic within the cells. This mixture makes it critical to demonstrate control over the manufacturing process. As a result of the size and complexity of bi­ologics, multiple methods are re­quired to detect the different types of potential in-process and degradation products. Control over the process can be demonstrated using both chro­matographic and electrophoretic techniques as well as assays demonstrat­ing that the desired activity of the bio­logic is maintained.

William Boomershine, PhD, Sen­ior Manager, Biologics, Alcami, ex­plains Alcami’s chromatographic/ electrophoretic techniques, such as in the case of charge state variants, which can arise during the fermenta­tion process in the form of various gly­cans containing differing sialic acids. Charge state variants can also arise as degradation products on stability. Deamidation of aspartic acid or glu­tamic acid residues to yield asparagine or glutamine residues change the overall charge of the pro­tein. Charge state variants can be sep­arated chromatographically using ion-exchange chromatography (IEX). The decision to use cation-exchange or anion-exchange chromatography will depend on the isoelectric point of the protein and the overall charge of the protein in the formulation buffer. Charge state variants can also be separated using electrophoresis. Tra­ditional iso-electric focusing and cap­illary iso-electric focusing can provide an orthogonal method to IEX for sep­aration of charge state variants.

Dr. Boomershine goes on to ex­plain how molecular weight variants can be in-process impurities formed during the purification or refolding steps of drug substance manufactur­ing, and can be degradation prod­ucts, forming in the drug product on stability. Size exclusion chromatogra­phy can monitor both dimers and higher order aggregates, as well as smaller impurities in buffers that can closely mimic physiological buffers. Electrophoresis – both traditional SDS-PAGE and capillary – can separate impurities based on molecular weight. Non-reduced SDS is utilized to moni­tor molecular weight variants that use disulfide bonds to form dimers and other higher order aggregates while reduced SDS can detect molecular weight variants of the light and heavy chains from monoclonal antibodies.

“While the above chromato­graphic and electrophoretic tech­niques can be used to look at charge state and molecular weight variants of the biologic as a whole, these techniques may not easily see specific, in­dividual changes that can impact activity,” he says.

Detecting and quantitating local changes require techniques with higher resolution, such as a peptide map. The individual peptides from a peptide map can be more easily separated from each other using re­versed-phase HPLC. “Chromatogra­phy for a peptide map can be optimized to target the separation and quantitation of a specific process impurity or degradation product, such as methionine oxidation, N-terminal vari­ants, or deamidation of a specific residue,” he says.

Article Details

Drug Development & Delivery
Issue: January/February 2021

REFERENCE

1. Pharmaceutical Analytical Testing Outsourcing Market Size, Share & Trends Analysis Report By Service (Bioanalytical, Stability, Method Development & Validation), By Region, And Segment Forecasts, 2020 – 2027), February 2020, https://www.grandviewresearch.com/industry-analysis/pharmaceutical-analytical-testing-outsourcing-market.

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