Development toward rapid and efficient screening for high performance hydrolysate lots in a recombinant monoclonal antibody manufacturing process

Abstract

Plant-derived hydrolysates are widely used in mammalian cell culture media to increase yields of recombinant proteins and monoclonal antibodies (mAbs). However, these chemically varied and undefined raw materials can have negative impact on yield and/or product quality in large-scale cell culture processes. Traditional methods that rely on fractionation of hydrolysates yielded little success in improving hydrolysate quality. We took a holistic approach to develop an efficient and reliable method to screen intact soy hydrolysate lots for commercial recombinant mAb manufacturing. Combined high-resolution (1) H nuclear magnetic resonance (NMR) spectroscopy and partial least squares (PLS) analysis led to a prediction model between product titer and NMR fingerprinting of soy hydrolysate with cross-validated correlation coefficient R(2) of 0.87 and root-mean-squared-error of cross-validation RMSECV% of 11.2%. This approach screens for high performance hydrolysate lots, therefore ensuring process consistency and product quality in the mAb manufacturing process. Furthermore, PLS analysis was successful in discerning multiple markers (DL-lactate, soy saccharides, citrate and succinate) among hydrolysate components that positively and negatively correlate with titer. Interestingly, these markers correlate to the metabolic characteristics of some strains of taxonomically diverse lactic acid bacteria (LAB). Thus our findings indicate that LAB strains may exist during hydrolysate manufacturing steps and their biochemical activities may attribute to the titer enhancement effect of soy hydrolysates.

Effect of copper variation in yeast hydrolysate on C‐terminal lysine levels of a monoclonal antibody

“…At both large‐ and small‐scale, lactate, pH, and mAb‐K levels were correlated with copper levels measured in yeast hydrolysate. It is well known that hydrolysates are inherently complex and variable but are often tolerated in cell culture processes for their benefits of increasing cell growth and product yield . However, this study documents the impact of a trace metal variation from a hydrolysate source on the consistency of a product attribute.…”

Serum and Protein Free Media

“…This type of analysis has proved difficult because of the variability of hydrolysate preparations from one batch to another. This has lead to the development of a predictive model as a screening tool for high performance hydrolysate lots (Luo and Pierce 2012). Without necessarily defining the composition of the hydrolysate, NMR fingerprinting can identify a pattern which could predict process consistency and product quality.…”

Influence of protein and carbohydrate contents of soy protein hydrolysates on cell density and IgG production in animal cell cultures

“…Thirdly, it has been proposed that microbial fermentation could occur during the enzymatic hydrolysis. 3 To test for the presence of carbohydrase activities, SSPS was incubated with Phyzyme and Bromelain. SSPS is a water-soluble polysaccharide extracted and refined from soybean and is composed of galactose, arabinose, rhamnose, xylose, fucose, glucose, and galacturonic acid.…”

“…In addition, the hydrolysates contain a large variety of different compounds (proteinaceous material, carbohydrates, minerals, lipids, isoflavones, saponins, and organic acids), which can affect cell culture in different ways. The minor compounds, such as sucrose, 3 raffinose, 3 stachyose, 3 citrate, 3 cytidine, and recombinant protein production. 4 In contrast to minor compounds, the effect of the variation in the contents of different compound classes, i.e.…”