Search

Data included are related to the research article “Isolation of biofunctional bovine immunoglobulin G from milk- and colostral whey with mixed-mode chromatography at lab and pilot scale” (Heidebrecht et al., 2018) [1]. Data show individual bovine whey proteins in flow-through and elution fractions using different chromatographic resins as well as different binding and elution conditions. The relevant analytical methods for individual protein detection were SDS-PAGE and reversed phase- high performance liquid chromatography. The focus of the data is on the two mixed mode materials MEP HyperCel™ and Capto™-multimodal chromatography. Resins were used individually, in series and at different scale. Data provide information at which binding and elution conditions it is possible to isolate bovine IgG from milk and colostral whey and at which purity.

Clostridioides difficile infection (CDI) often develops after pretreatment with antibiotics, which can lead to damage of the intestinal microbiome. The approach of this study was to use specific polyclonal antibodies isolated from the milk of immunized cows to treat CDI, in contrast to the standard application of nonspecific antibiotics. To gain a deeper understanding of the role of the microbiome in the treatment of CDI with bovine antibodies, stool and intestinal fluid samples of hamsters were collected in large quantities from various treatments (>400 samples). The results show that the regeneration of the microbiome instantly begins with the start of the antibody treatment, in contrast to the Vancomycin-treated group where the diversity decreased significantly during the treatment duration. All antibody-treated hamsters that survived the initial phase also survived the entire study period. The results also show that the regeneration of the microbiome was not an antibody-induced regeneration, but a natural regeneration that occurred because no microbiota-inactivating substances were administered. In conclusion, the treatment with bovine antibodies is a functional therapy for both the acute treatment and the prevention of recurrence in hamsters and could meet the urgent need for CDI treatment alternatives in humans.

Data are related to the research article “Fractionation of casein micelles and immunoglobulins by microfiltration in diafiltration mode Study of the transmission and yield of IgG, IgA and IgM” [1]. The data show the transmission and yield of the individual whey proteins α-Lactalbumin (α-La), β -Lactoglobulin (β -Lg), blood serum albumin (BSA), lactoferrin (LF), lactoperoxidase (LPO) and the immunoglobulins IgG, IgA, IgM during microfiltration (0.14 μm) performed in diafiltration mode at 50 °C with different applied transmembrane pressures (0.6-3 bar). The data provide information on the decrease of the respective proteins in the microfiltration retentate and their increase in the UF retentate. The relevant analytical methods for the individual protein detection were performed by reversed phase high performance liquid chromatography and ELISA. The isoelectric point of IgG and IgM was measured with the Zetasizer Nano ZS. Keywords: Immunoglobulins, Microfiltration, IgG

Existing works on the influence of spatial effects on flux and permeation of proteins in microfiltration (MF) have focused on ceramic membranes. There is little information on spiral-wound membranes (SWMs). Since the inner core of a SWM is practically inaccessible by non-destructive techniques, three different prototypes were constructed in this study to optimize suitability for the investigation of spatial effects on filtration performance. To measure the pressure drop, shortened SWMs 0.25, 0.50, and 0.75 times the length of a standard industrial SWM (0.96 m) were designed. Second, a sectioned membrane (0.96 m) with separated compartments on the permeate side was constructed to analyze spatial effects on flux and protein permeation along the flow path of a SWM. Three different features characterized this sectioned module: sectioned permeate pockets, a sectioned permeate collection tube, and sectioned permeate drain and measurement systems. Crossflow filtration experiments showed that these modifications did not alter the filtration performance compared to an unmodified control SWM. Thus, it can be applied to assess spatially-resolved filtration performance in SWMs. The third prototype designed was a test cell with accessible flat sheet membranes and spacer material, as in SWMs. The flow path in this test cell was designed to match the characteristics of the channels between the membrane sheets in a standard SWM as closely as possible. The flow path length and the combination of membrane material and spacer architecture were the same as in the control SWM. This test cell was designed to assess the effects of length and processing conditions on the formation of a deposit layer. The combined results of these test modules can yield new insights into the spatial distribution of flux, permeation of target components, and deposit formation.

Protein fractionation by means of microfiltration (MF) is significantly affected by fouling, especially when spiral-wound membranes (SWMs) are used. We investigated the influence of the mode of transmembrane pressure (ΔpTM) increase to target level and the deposit layer pressure history on the filtration performance during skim milk MF at temperatures of 10 °C and 50 °C. Two filtration protocols were established: No. 1: ΔpTM was set directly to various target values. No. 2: Starting from a low ΔpTM, we increased and subsequently decreased ΔpTM stepwise. The comparison of both protocols tested the effect of the mode of ΔpTM increase to target level. The latter protocol alone tested the effect of the deposit layer history with regard to the ΔpTM. As expected, flux and protein permeation were both found to be functions of the ΔpTM. Further, both measures were independent of the filtration protocol as long as ΔpTM was held at a constant level or, as part of protocol No. 2, ΔpTM was increased. Thus, we can state that the mode of ΔpTM increase to target level does not affect filtration performance in SWM. We found that after completion of a full cycle of stepping ΔpTM up from 0.5 bar to 3.0 bar and back down, flux and deposit layer resistance were not affected by the deposit layer history at 10 °C, but they were at 50 °C. Protein permeation, however, was lower for both 10 °C and 50 °C, when the ΔpTM cycle was completed. The processing history had a significant impact on filtration performance due to remaining structural compression effects in the deposited layer, which occur most notably at higher temperatures. Furthermore, temperatures of 50 °C lead to deposit layer aging, which is probably due to an enhanced crosslinking of particles in the deposit layer. Apart from that, we could show that fouling resistance does not directly correlate with protein permeation during skim milk MF using SWM.

Toxin-induced Clostridium difficile infection (CDI) is a major disease characterized by severe diarrhea and high morbidity rates. The aim with this study was to develop an alternative drug for the treatment of CDI. Cows were repeatedly immunized to establish specific immunoglobulin G and A titers against toxins A (TcdA) and B (TcdB) and against C. difficile cells in mature milk or colostrum. The effect of three different concentrations of anti-C. difficile whey protein isolates (anti-CD-WPI) and the standard of care antibiotic vancomycin were investigated in an animal model of CD infected hamsters (6 groups, with 10 hamsters each). WPI obtained from the milk of exactly the same cows pre-immunization and a vehicle group served as negative controls. The survival of hamsters receiving anti-CD-WPI was 50, 80 and 100% compared to 10 and 0% for the control groups, respectively. Vancomycin suppressed the growth of C. difficile and thus protected the hamsters at the time of administration, but 90% of these hamsters nevertheless died shortly after discontinuation of treatment. In contrast, the surviving hamsters of the anti-CD-WPI groups survived the entire study period, although they were treated for only 75 h. The specific antibodies not only inactivated the toxins for initial suppression of CDI, but also provoked the inhibition of C. difficile growth after discontinuation, thus preventing recurrence. Oral administration of anti-CD-WPI is a functional therapy of CDI in infected hamsters for both primary treatment and prevention of recurrence. Thus, anti-CD-WPI could address the urgent unmet medical need for treating and preventing recurrent CDI in humans.

The use of bioactive bovine milk immunoglobulins (Ig) has been found to be an alternative treatment for certain human gastrointestinal diseases. Some methodologies have been developed with bovine colostrum. These are considered in laboratory scale and are bound to high cost and limited availability of the raw material. The main challenge remains in obtaining high amounts of active IgG from an available source as mature cow milk by the means of industrial processes. Microfiltration (MF) was chosen as a process variant, which enables a gentle and effective concentration of the Ig fractions (ca. 0.06% in raw milk) while reducing casein and lactose at the same time. Different microfiltration membranes (ceramic standard and gradient), pore sizes (0.14–0.8 µm), transmembrane pressures (0.5–2.5 bar), and temperatures (10, 50 °C) were investigated. The transmission of immunoglobulin G (IgG) and casein during the filtration of raw skim milk (