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Bovine colostrum is both a primary source of nutrients and immunological support for the newborn calf. The understanding of the level of therapeutic proteins (lactoferrin and IgG) is important for the bovine colostrum dosing and standardization for human consumption.
Colostrum is a complex biological fluid produced by mammals immediately after parturition. It meets all the nutritional requirements for neonates as a good source of macro- and micronutrients, bioactive peptides, and growth factors. Bovine colostrum is also a potential source of nutrition and bioactive because of its rich protein content that includes immunoglobulin G (IgG) and lactoferrin. However, the level of lactoferrin and IgG in bovine colostrum changes markedly during the lactation period. Therefore, monitoring the concentration of IgG and lactoferrin for the use of bovine colostrum as a protein source is an important question to study. Methods in this article describe how to determine protein content, as well as specific concentrations of lactoferrin and IgG. These methods include the following steps: Isolation of bovine colostrum proteins, Determination of protein concentration via Bicinchoninic acid assay (BCA), Visualization of proteins via SDS-PAGE, Determination of lactoferrin, and IgG concentration using an ELISA Assay.
Colostrum is the initial secretion of the mammary gland produced by mammals shortly after parturition. Colostrum is rich in macro- and micronutrients, antimicrobial peptides, and growth factors1,2,3,4. The composition varies gradually over time through the transition to mature milk5,6,7 but most significantly within 24 h after parturition8. The composition of colostrum is also influenced by maternal factors, including age, pa....
Colostrum samples were collected for 6 days after calving in the noon over the period July-August, from 28 Holstein dairy cows from Uluova Milk Trading Company in Çanakkale, Turkey, and deep-frozen. The samples collected on the same day were pooled according to the day of each sample and analyzed for their total protein, lactoferrin, and IgG concentrations. All samples were assayed in duplicate.
1. Sample preparation
Following the protocol, the bovine colostrum samples were analyzed to determine protein, lactoferrin, and IgG concentration. The results of protein, lactoferrin, and IgG analyses of bovine colostrum are shown in Table 4.
Table 4: Concentration of protein, lactoferrin, and IgG of bovine colostrum.
This study provides information about considerable changes in the protein, lactoferrin, and IgG concentrations in colostrum throughout the transition to mature milk. Detection of changes in the lactoferrin and IgG concentration was carried out by sandwich ELISA, and total protein concentration was analyzed by the BCA assay. Results indicate that early colostrum has the highest protein, lactoferrin, and IgG concentration, that subsequently decreased over the next 3 days. Accurate measurements of these proteins are relevan.......
This study is supported by Uluova Süt Ticaret A.Ş (Uluova Milk Trading Co.). RMD and BMH are employees of Evolve BioSystems, a company focused on restoring the infant microbiome.
....Name | Company | Catalog Number | Comments |
10X Running Buffer (Tris-Glycine-SDS) | ClearBand | TGS10 | SDS-Page analysis |
2-mercaptoethanol | gibco | 31350-010 | SDS-Page analysis |
Acetic Acid GLACIAL | Isolab | 901,013,2500 | SDS-Page analysis |
Bovine IgG ELISA Kit | Aviva Systems Biology | OKIA00005 | Determination of IgG concentration |
Bovine LF / LTF / Lactoferrin ELISA Kit | LSBio Lifespan Biosciences | LS-F4884 | Determinaton of lactoferrin concentration |
Coomassie Brillant Blue R 250 | amresco | 0472-25G | SDS-Page analysis |
Hydrochloric Acid Fuming 37% | Isolab | 932,103,2501 | SDS-Page analysis |
Isopropanol | Isolab | 961,023,2500 | SDS-Page analysis |
Laemmli Sample Buffer (2X) | ClearBand | LSB-2x | SDS-Page analysis |
Methanol | Isolab | 947,046,2500 | SDS-Page analysis |
PageRuler Plus Prestained Protein Ladder 10 to 250 | Thermo Scientific | 26619 | SDS-Page analysis |
Pierce BCA Protein Assay Kit | Thermo Scientific | 23225 | Determination of protein concentration |
Sodium dodecyl sulfate (SDS) | BioShop | SDS001.500 | SDS-Page analysis |
SureCast Acrylamide Solution 40% (w/v) | Invitrogen | HC2040 | SDS-Page analysis |
SureCast Ammonium persulfate (APS) | Thermo Scientific | 17874 | SDS-Page analysis |
SureCast Tetramethylethylenediamine (TEMED) | Invitrogen | HC2006 | SDS-Page analysis |
TECAN Infinite M200 Plate Reader | Tecan | 30035094 | Measurement of absorbance |
Tris base | BioShop | TRS001.1 | SDS-Page analysis |
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