000 03523nam a2200193Ia 4500
005 20151005141019.0
008 150525s2011 xx 000 0 und d
041 _aeng
082 _a1330,T
100 _aSidra Ashraf
_97858
110 _cDr. Abu Saeed Hashmi
_95243
245 _aIsolation And Characterization Of Collagen Type Ii From Poultry Trachea
260 _c2011
502 _aThis project was designed to use poultry waste to isolate and characterize collagen type II from its trachea. Collagen type II is being used along with condroitin sulfate and glucosamine for the treatment of osteoarthritis and is also available as a neutraceutical product in the market. For project purpose, trachea of slaughtered broiler birds were collected from the market and after removing adhering tissue and debris, it was then washed thoroughly first with distilled water and then with deionized water. Tracheal cartilage was then cut into small pieces and defattened with chloroform: methanol (2: 1 v/v) solution. After this, the cut pieces were properly cleaned with deionized water. 0.5% Pepsin solution in 0.5 M acetic acid was prepared. Cartilage was then hydrolyzed by the already prepared 0.5 % pepsin (in 0.5 M acetic acid) at 4 ° C for 48 hours. The extract was then separated from the tracheal pieces and the viscous solution obtained was centrifuged at 12000 rpm for 1 hr at 4 "c. Now the collagen was expected to be in the supernatant which was salted out by adding NaCI to a final concentration of 2.5M and kept for almost 12-16 hrs. This collagen was again centrifuged at 12000 rpm for 1 hr at 4 C. The obtained collagen pallet was redissolved in 0.5 M acetic acid and then it was dialyzed against 0.1 M acetic acid followed by dialysis with distilled water. The sample after dialysis was put in petri dishes and kept in freezer for overnight to let it be prepared for lyophilization. The frozen collagen sample was then lyophilized. After lyophilization, the sample gave an appearance of a white mesh. This sample was reconstituted in PBS with pH 8 to run it on SDS-PAGE. The procedure of SDS-PAGE in non reducing conditions was adopted for the characterization of collagen type II in the sample. The description of results of SDS-PAGE is given below: Lane M contains protein markers of different molecular weight. Lane 1, 2 and 3 contains samples at different steps of the whole procedure showing clear bands of collagen type II. Lane 4 contains lyophilized sample of collagen type II showing the thickest band (alpha chain of collagen type II). In this research, poultry waste has been used for making health improving product. As in our country poultry is used in bulk quantity so if its waste might be used in any medicinal product then it might not only be useful but also economical for such a developing country as ours. Another thing is that as this collagen Type II has been extracted from poultry trachea, it shows that tracheal cartilage is a rich source of such collagen type. Collagen Type II is used in the cure of arthritis especially rheumatoid arthritis so through this research, it has been made clear that poultry waste can be utilized in a positive way in medicinal industry and also that collagen Type II acts as an effective neutraceutical.
650 _aInstitute of Biochemistry & Biotechnology
_95022
700 _aDr. Sualeha Riffat
_96434
700 _aZahid Mushtaq
_97859
942 _cTH
999 _c3046
_d3046