Year: 2022 | Month: April | Volume 12 | Issue 2
Assay of BLf using Sandwich ELISA and its Comparative Study of Holstein-Friesian Crossbred and Sahiwal with Poda Thurpu Cow Milk
Mohammed Shaz Murtuza
Ashok Kumar Devarasetti
Akkaldevi Jayasri
B.D.P. Kalakumar
Lavudya Naveen
DOI:10.30954/2277-940X.02.2022.22
Abstract:
Bovine lactoferrin (BLf) is a multifunctional bioactive protein found in high concentration in milk. The present study was done to evaluate, isolate and compare the quantity of BLf present in the milk of exotic cross bred and indigenous breeds of cow. Milk samples were collected from exotic HF cross bred and indigenous breeds like Sahiwal and Poda thurpu. The quantification of BLf was done by using sandwich enzyme-linked Immunosorbent assay (ELISA). The processed milk samples of each of three breed were then subjected to cation exchange column chromatography using Sephadex C-50 resin. The eluted fractions obtained from the chromatography were used to run sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS-PAGE) along with a protein marker to confirm the identity of BLf. The estimated BLf values (μg/mL) from ELISA were found as 306.47+8.91, 369.06+14.04 and 403.9+20.10 in HF cross bred, Sahiwal and Poda thurpu cow breed. Further electrophoresis results showed a clear band was formed at 80KDa approximately when compared with the standard protein marker with a more prominent band in indigenous breeds than HF cross bred where a thin band was seen.
Highlights
- Lactoferrin (Lf) is a iron binding protein which shows various bioactive functions.
- BLf content was highest in indigenous breed cow milk than in exotic cross bred cow milk.
- Among indigenous breeds, BLf concentration was found to be higher in Poda thurpu cow milk followed by Sahiwal cow milk.
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