Sidra Ashraf
Isolation And Characterization Of Collagen Type Ii From Poultry Trachea - 2011
This 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.
Institute of Biochemistry & Biotechnology
1330,T
Isolation And Characterization Of Collagen Type Ii From Poultry Trachea - 2011
This 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.
Institute of Biochemistry & Biotechnology
1330,T