Data concerning the chromatographic isolation of bovine IgG from milk- and colostral whey

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.


a b s t r a c t
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.
& 2018 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

Subject area
Chemistry, biology More specific subject area Isolation of bovine antibodies Type of data Graphs, figure How data was acquired ÄKTApurifier 100 UPC, ÄKTApilot, Bio-Rad process chromatography station, Zetasizer Nano ZS, SDS-PAGE, reversed phase-high performance liquid chromatography Data format Analyzed Experimental factors Fat (centrifugation), casein (microfiltration), lactose/minerals (ultrafiltration) removed from raw colostrum or milk to obtain whey Experimental features Determination of individual whey proteins in flow-through and elution fraction using different binding and elution conditions Data source location Technical University of Munich (Freising) and Ludwig-Maximilian-University (Munich) Germany

Data accessibility
With this article Related research article Data is provided as additional material directly related to the article [1].

Value of the data
Isolation of bovine IgG from milk and colostral whey using the two mixed mode materials MEP HyperCel ™ and Capto ™ at different scale.
Data deliver information about adsorption and desorption of bovine IgG at various different binding and elution conditions (pH, ionic strengths, buffer).
Data are suitable for the setup of a chromatographic isolation process to obtain therapeutic amounts of isolated bovine IgG.

Data
The data show how to isolate bovine IgG from milk or colostral whey with two mixed mode materials and which conditions to use at different scale.
Figs. 1 and 2 show the binding mechanism of the two resins MEP HyperCel ™ (MEP) and Capto ™ -MMC (MMC). In order to determine the optimal elution pH of IgG from the MEP column a pH gradient elution was carried out (Fig. 3). To better detect the individual proteins the flow-through and elution samples, proteins were visualized with different methods on different SDS-PAGE gels (Fig. 4 reducing and non-reducing (Fig. 5) SDS-PAGE stained-free gel with UV protein visualization, Fig. 6 reducing SDS-gel with coomassie protein visualization). Fig. 7 shows binding and elution of IgG at increased pH (pH 9 instead of 7.5) during the binding phase at the MMC column. Fig. 8 shows the serial application of the flow through of the MMC column to the MEP column at increased ionic strength during the binding phase (0.25 mol L À 1 NaCl). Fig. 9 shows the introduction of an elution step at pH 6 and a visible band that represents major whey protein β-lactoglobulin at the relevant lane.

Experimental design, materials, and methods
The aim was to develop a novel and scalable process for the isolation of bovine IgG from colostral and milk whey, respectivley, with high purity and recovery. The preparation of the samples, the equipment and analytical methods for analysis are described in detail in [1].    toperoxidase (LPO). The data shown in Figs. 3-9 were carried out on an ÄKTApurifier 100 UPC (GE Healthcare, Uppsala, Sweden) with a tunable flow rate up to 100 mL min À 1 . The remaining isolation data in Figs. 10-20 were executed by an ÄKTApilot (GE Healthcare, Uppsala, Sweden) system with a flow rate range from 4 to 400 mL min À 1 for the MMC resin respectively with an Bio-Rad process chromatography station (Bio-Rad Laboratories GmbH, Munich, Germany) with a flow rate range from 83 to 2000 mL min À 1 for the MEP column. The corresponding buffers are summarized in Table 1.

Particle size measurement during desalting
In order to monitor aggregation of IgG during desalting, the particle size was measured by dynamic light scattering using the Zetasizer Nano ZS (Malvern Instruments, Malvern, UK). At different diafiltration steps samples with constant IgG content were filtered using a syringe filter of 0.45 mm (Chromafil RC-45/25 Macherey-Nagel, Dueren, Germany). After a 5 min equilibration phase each sample was measured in duplicate at 20°C. Each of the two runs consisted of 10 individual runs of 60 s (Fig. 21).