Production, Purification And Evaluation Of Anti Tetanus Serum (Record no. 3126)

000 -LEADER
fixed length control field 02470nam a2200193Ia 4500
005 - DATE AND TIME OF LATEST TRANSACTION
control field 20151005154946.0
008 - FIXED-LENGTH DATA ELEMENTS--GENERAL INFORMATION
fixed length control field 150525s2012 xx 000 0 und d
041 ## - LANGUAGE CODE
Language code of text/sound track or separate title eng
082 ## - DEWEY DECIMAL CLASSIFICATION NUMBER
Classification number 1420,T
100 ## - MAIN ENTRY--AUTHOR NAME
Personal name Mian Muhammad Khubaib Sattar
110 ## - MAIN ENTRY--CORPORATE NAME
Location of meeting Prof. Dr. Tahir Yaqub
245 ## - TITLE STATEMENT
Title Production, Purification And Evaluation Of Anti Tetanus Serum
260 ## - PUBLICATION, DISTRIBUTION, ETC. (IMPRINT)
Year of publication 2012
502 ## - DISSERTATION NOTE
Dissertation note To produce anti-tetanus serum, ten female sheep of about 3 years of age are immunized with commercially available tetanus toxoid vaccine (Imatet™, Amson vaccines and Pharma) for eighteen weeks to these experimental animals with two weeks interval. Serum samples of all the sheep were also collected fortnightly and stored at -20 ºC.
ELISA was performed to determine the antibody (IgG) titer of all the test samples. Out of 90 samples, 20 samples showed tetanus antibody (IgG) titer of 100.8 I.U. or more, while 8 samples presented tetanus antibody (IgG) titer of 160.9 IU or more. Out of these 8 samples, 3 samples had tetanus antibody (IgG) titer of 190.9 I.U. or more. Maximum tetanus antibody (IgG) titer was 195.4 I.U. at day 120.
Three samples exhibiting maximum antibody titer (190.9 I.U., 195 I.U. and 195.4 I.U.) were mixed in equal quantities for purification of Immunoglobulins (IgG). A volume of 15 mL of aggregated serum samples was mixed with 15 ml of saturated ammonium sulfate having final concentration of 45 % in the mixture which is continuously stirred at room temperature for 1 hour. Mixture was centrifuged at 10000 rpm for 30 minutes in refrigerated centrifuge machine and dialyzed against 10 changes of PBS at 4 ºC. Desalting is checked with BaCl2 solution.
The purified tetanus immunoglobulins (IgG) were treated with papain to produce Fab Fragments. Then the protein content of the purified tetanus immunoglobulins and Fab fragments was estimated with Bradford protein Assay. BSA standard curve was used to produce a regression equation [Y (OD Value) = 0.218 + 0.033 X (Protein Concentration)] which was used for calculation of the protein contents of the samples.
The purified tetanus immunoglobulins and Fab fragments were tested for purity with SDS-PAGE analysis. Then in vivo toxin neutralization test was performed in mice to check the tetanus toxin neutralization ability of the sera produced.
650 ## - SUBJECT ADDED ENTRY--TOPICAL TERM
Topical Term Department of Microbiology
700 ## - ADDED ENTRY--PERSONAL NAME
Personal name Dr.
700 ## - ADDED ENTRY--PERSONAL NAME
Personal name Mr. Muhammad Zubair Shabbir
942 ## - ADDED ENTRY ELEMENTS (KOHA)
Koha item type Thesis
Holdings
Damaged status Collection code Permanent Location Current Location Shelving location Date acquired Full call number Accession Number Koha item type
  Veterinary Science UVAS Library UVAS Library Thesis Section 2015-05-29 1420,T 1420,T Thesis


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