Production of Extracellular Protein a from a Methicillin-Resistant Strain of Staphylococcus Aureus
Protein A was produced extra cellularly by methicillinresistant Staphylococcus aureus, strain A676. A competitive- ELISA technique based on a competitive pinding between unlabelled protein A and conjugated protein A was employed to measure the concentration of protein A. A study was also conduct...
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| Main Author: | |
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| Format: | Thesis |
| Language: | English English |
| Published: |
1994
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| Subjects: | |
| Online Access: | http://psasir.upm.edu.my/id/eprint/8361/1/FSMB_1994_2_A.pdf |
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| Summary: | Protein A was produced extra cellularly by methicillinresistant
Staphylococcus aureus, strain A676. A competitive-
ELISA technique based on a competitive pinding between unlabelled protein A and conjugated protein A was employed to
measure the concentration of protein A. A study was also
conducted to develop this technique. Rabbit IgG was first purified before it was used to coat a
microtiterplate. IgG purified using ammonium sulphate
precipitation in combination with DEAE-cellulose chromatography
gave purer IgG than the caprylic acid method combined with
saturated ammonium sulphate or single-step DEAE-cellulose ion
exchange chromatography. The molecular weight of the purified
IgG was 140,000 dalton based on SDS gelelectrophoresis. The
minimum detectable amount of protein A was in the range of 0-20
ng/ml. The optimum concentration of IgG required for coating
was 2pg/ml and the incubation time for the substrate was 20 minutes. |
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