Journal of Animal Research
  • Year: 2022
  • 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

  • Author:
  • Mohammed Shaz Murtuza1,*, Ashok Kumar Devarasetti2, Akkaldevi Jayasri1, B.D.P. Kalakumar3, Lavudya Naveen1
  • Total Page Count: 5
  • Published Online: Jul 14, 2023
  • Page Number: 305 to 309

1Department of Veterinary Biochemistry, College of Veterinary Science, Rajendranagar, PV Narasimha Rao Telangana Veterinary University, Hyderabad, India

2Department of Veterinary Biochemistry, College of Veterinary Science, Mamnoor, PV Narasimha Rao Telangana Veterinary University, Hyderabad, India

3Department of Veterinary Pharmacology and Toxicology, College of Veterinary Science, Mamnoor, PV Narasimha Rao Telangana Veterinary University, Hyderabad, India

*Corresponding author: MS Murtuza; E-mail: msmurtuza@gmail.com

Online Published on 14 July, 2023.

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.

• 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.

Keywords

Bovine lactoferrin, Poda thurpu, ELISA, Sephadex C-50, SDS-PAGE