DEVELOPMENT OF INDIRECT SANDWICH ELISA METHOD FOR DETERMINATION OF RECOMBINANT DARBEPOETIN
Darbepoetin (DPO) is a glycoprotein hormone by stimulating colony forming units erythroid (CFU-E) to increase the production of erythrocytes. DPO has 5 Nlinked chains and 1 O-linked chain with the potential maximum number of sialic is 22 molecules. An extra 2 N chains gives DPO greater metabolic s...
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| Format: | Theses |
| Language: | Indonesia |
| Online Access: | https://digilib.itb.ac.id/gdl/view/51580 |
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| Institution: | Institut Teknologi Bandung |
| Language: | Indonesia |
| Summary: | Darbepoetin (DPO) is a glycoprotein hormone by stimulating colony forming
units erythroid (CFU-E) to increase the production of erythrocytes. DPO has 5 Nlinked chains and 1 O-linked chain with the potential maximum number of sialic
is 22 molecules. An extra 2 N chains gives DPO greater metabolic stability in vivo
compared to the previous generation. Detection of DPO concentrations using
ELISA has previously been used commercial kits but the cost of commercial kits is
quite expensive and needs to be done developing and validating indirect-sandwich
ELISA methods in-house to measure DPO levels produced by Chinese Hamster
Ovary (CHO) cell scale research and development. Optimal coating antibody
results dilution at 1:6000. An indirect-sandwich ELISA in-house method was
designed that meets all requirements to fulfill this goal. The method has a linear
range 2.29-73.40 ?g/mL with coefficient correlation (R)=0.9959. Accuracy is
obtained in the range of 100-103%. Intra-day and inter-day precision is 0.46-
8.44% and 3.44-9.33%. LOD and LOQ were 0.2767 ?g/mL and 0.9224 ?g/mL,
respectively. The cost analysis showed that the ELISA method in-house assay is
cheaper than the existing commercial sandwich ELISA kits.
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