Characterization of soy protein hydrolysates and influence of its iron content on monoclonal antibody production by a murine hybridoma cell line

Abstract

A challenging aspect with the use of protein hydrolysates in commercial manufacturing processes of recombinant therapeutic proteins is their impacts on the protein production due to a lack of understanding of batch-to-batch variability. Soy hydrolysates variability and its impact on fed-batch production of a recombinant monoclonal antibody (mAb) expressed in Sp2/0 cells were studied using 37 batches from the same vendor. The batch-to-batch variability of soy hydrolysates impacted cell growth, titer and product quality. Physicochemical characterization of batches confirmed that soy hydrolysates are mainly a source of amino acids and peptides containing lower amounts of other components such as carbohydrates and chemical elements in cell culture media. Soy hydrolysates composition of different batches was consistent except for trace elements. Statistical analyses identified iron as a potential marker of a poor process performance. To verify this correlation, two forms of iron, ferric ammonium citrate and ferrous sulfate, were added to a batch of soy hydrolysates associated to a low level of iron during cell culture. Both forms of iron reduced significantly cell growth, mAb titer and increased level of the acidic charge variants of the mAb. Consequently, trace element composition of soy hydrolysates or of all incoming raw materials might lead to significant impacts on process performance and product quality and therefore need to be tightly controlled.

Cottonseed hydrolysate supplementation alters metabolic and proteomics responses in Chinese hamster ovary cell cultures

“…These spectroscopic techniques, including Raman spectroscopy, fluorescence spectroscopy, and near infrared spectroscopy, when combined with statistical methods, have been shown to possess the capability of separating the hydrolysate lots based on their performance early on in the raw material development stage. [ 20,21 ] These characterization techniques can significantly reduce the variability that hydrolysates are believed to impart to the process and make them viable medium additives similar to their proprietary CD counterparts, whose formulations are unknown in most cases.…”

Effect of Conventional Humid–Dry Heating through the Maillard Reaction on Chemical Changes and Enhancement of In Vitro Bioactivities from Soy Protein Isolate Hydrolysate–Yeast Cell Extract Conjugates

“…The yeast lysis conditions, including pH (5.0-6.5) and temperature (50-70 • C), were carefully controlled to obtain the desired extract [16,22]. Regarding soy protein hydrolysates, they also contain a carbohydrate portion, with sucrose and stachyose as the primary components, along with minor proportions of raffinose, fructose, glucose, etc., depending on batch production [43]. The total protein content of SPIH and YCE showed both extracts containing protein quantities of 58.31 g/100 g and 79.57 g/100 g, respectively.…”

“…On the other hand, the magenta smear bands of glycoprotein became slightly more intense after 15 days of the procedure, possibly implying the establishment of sugar-peptide compounds. According to Conti et al [51] and Djemal et al [43], soy protein hydrolysates exhibited peptides with molecular masses ranging from 0.5 to 10 kDa, while yeast oligopeptides were characterized by a molecular weight within the range of 2-3 kDa [16]. The increase in peptide molecular weight through humid-dry heating incubation could be attributed to glycosylation between peptides and saccharides, resulting in the formation of conjugated compounds, as evidenced by the presence of polydisperse bands in the middle to the top of the SDS-PAGE gel, along with a broader distribution of larger peptide chain masses [6,52].…”

“…Commercial soy protein hydrolysates were composed of peptides/amino acids, carbohydrates, vitamins, lipids, and minerals. The protein content exhibited variability within a range of 56-58%, influenced by batch-to-batch differences and soybean cultivation [43,44]. Yeast extract primarily contained umami-flavor amino acids such as Glu, Ala, Gly, and Asp, along with various other types of amino acids like Ser, Val, and Pro depending on the variety of yeasts.…”