• KSBB Journal
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• v.18 no.4
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• pp.255-260
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• 2003

In vivo bioassays for biological medicines have been considered final resort to unequivocally assess the biological activities for them because there are some cases in which the biological activities obtained from in vivo bioassay and in vitro bioassay quite differ each other. The in vivo biological activity of EPO depends on its sialic acid contents which confer microheterogeneity-isoforms to this protein. We have devise a method which consists of a in vitro bioassay using BaF3 cell line and a capillary zone electrophoresis (CZE) for the measurement of the EPO isoform distribution. The biological activity of EPO obtained using in vitro bioassay with BaF3 cell line showed good correlation (C.V.(％) 7.34, 5.85, 8,16, 8.08, 8.8) to EPO content measured either spectrophotometric assay (A280 0.1 ％ ＝0.743) or radio immunoassay. The assay validation results of in vitro bioassay with 3 lot of in house EPO showed good results to EPO content measured either in vivo assay or radio immunoassay. and also showed good results the robustness of our method in terms of precision, accuracy, repeatability. The isoform distribution for EPO-BRP (1 : 1 mixture of epoetin-${\alpha}$ and epoetin-${\beta}$, European Pharmacopoeia) by CZE method resulted in isoform 2 through isoform 8. The major peaks in electrophoregram were composed of isoform 3 through 7. Our recombinant EPO (epoetin-${\alpha}$) having equivalent in vivo biological activity showed the isoform distribution of isoform 3 through 9. The major peaks consisted of isoform 4 through 8. The peak area of isoform 4 was always smaller than that of isoform 5. The preparations of recombinant epoetin-${\alpha}$ with lower in vivo biological activity than EPO-BRP showed the isoform 2 through 8 in their electrophoregrams whose major peaks consisted of the isoform 3 through 7. The peak area of isoform 4 was larger than that of isoform 5.

• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.747-748
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• 2003

• Proceedings of the Korean Society of Toxicology Conference
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• 2001.10a
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• pp.201-201
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• 2001

Transforming growth factor- $\beta$ (TGF- $\beta$), a hormonally active polypeptide found in normal and transformed tissue, is a potent regulator of cell growth and differentiation. In this study, we wished to establish an in vitro bioassay system to seek the most sensitive method that can measure TGF- $\beta$ activity.(omitted)

• Journal of the Korean Society of Food Science and Nutrition
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• v.14 no.2
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• pp.202-213
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• 1985

The protein nutritional quality of foods has become an important factor to food processors with the advent of nutritional labeling regulations for foods. Then, as is true today, the officially approved assay for protein nutritional quality was the rat based protein efficiency ratio(PER) bioassay. The PER bioassay requires a minimum of 28 days to performe, and is therefore not applicable to routine quality assurance use by the food industry. Within the past ten years there has been a research emphasis placed on the development of rapid, inexpensive, biological and/or chemical based assays for protein nutritional quality. It was hoped that if a rapid assay could be developed and thoroughly tested, it could be used in lieu of the PER bioassay in the day-to-day quality assurance screening of food ingredients and products. The rapid assays developed in the hope of attaining this goal have been based on microorganisms, proteolytic enzymes, and amino acid profiles, as well as combinations of the above. In this review, it will be described and briefly discussed many of procedures which had contributed conceptually as well as practically to the development of in vitro methods for the evaluation of protein quality. Special emphasis will be placed on the C-PER(computed protein efficiency ratio) assay which combines data from in vitro protease digestion and amino acid composition to predict protein nutritional quality designed by Satterlee et al. (1980), and the DC-PER(discriminant computed PER) which is a method of estimating protein quality based on rat assay and in vitro digestibility obtained using solely essential amino acid data will be also introduced.

• Journal of Plant Biotechnology
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• v.40 no.1
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• pp.43-48
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• 2013

Oreorchis coreana Finet is threatened globally by over-collection from its natural habitats for horticultural purposes. Its rarity in nature makes this plant one of the most endangered species in Korea. In this study, we investigated the effects of sodium hypochlorite (NaOCl) on orchid seed viability and seed germination. An in vitro bioassay swelling test using immature seeds was compared with a standard chemical procedure using triphenyl tetrazolium chloride (TTC) to test seed viability. In general, the bioassay was more appropriate for estimating embryo viability after a prolonged pre-treatment (more than 1 h) in 1% NaOCl, a surface sterilant often used to enhance germination of seeds of terrestrial plants. Therefore, an efficient method for investigating in vitro swelling of immature seeds is urgently needed. We established a method for determining the viability and swelling of O. coreana seeds via in vitro examination of immature seeds. Treatment of immature seeds with 1% NaOCl for 10 min greatly enhanced the extent of swelling of immature zygote embryos when compared to untreated seeds. These data obtained here appear to be comparable to viability and swelling that occurs in O. coreana seeds via asymbiotic germination.

• Archives of Pharmacal Research
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• v.25 no.6
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• pp.903-909
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• 2002

Research in the cytokine field has grown exponentially in recent years, and the validity of such studies relies heavily on the appropriate measurement of levels of cytokines in various biological samples. Transforming growth factor (TGF)-$\beta$, a hormonally active polypeptide found in normal and transformed tissue, is a potent regulator of cell growth and differentiation. The most widely used bioassay for TGF-$\beta$ is the inhibition of the proliferation of mink lung epithelial cells. Though detection of [$^3$H]thymidine incorporation is more sensitive than the MTT assay, it presents some disadvantages due to the safety and disposal problems associated with radioisotopes. In this study, we attempted to ascertain the experimental conditions which could be used for measuring the in vitro biological activity of TGF-$\beta$ in a safer and more sensitive way compared with the currently available methods. We compared the commonly used method, the MTT assay, to the XTT assay using different parameters including cell number, incubation time and the wave length used for detecting the product. We examined the anti-proliferative activities of TGF-$\beta$ in three different cell lines: Mv-1-Lu mink lung epithelial cells, MCF10A human breast epithelial cells and H-ras-transformed MCF10A cells. Herein, we present an experimental protocol which provides the most sensitive method of quantifying the biological activity of TGF-$\beta$, with a detection limit of as low as 10 pg/ml: Mv-1-Lu or H-ras MCF10A cells ($1{\times}10^5/well$) were incubated with TGF-$\beta$ at $37^{\circ}C$ in a humidified $CO_2$ incubator for 24 hr followed by XTT treatment and determination of absorbance at 450 or 490 nm. Our results may contribute to the establishment of an in vitro bioassay system, which could be used for the satisfactory quantitation of TGF-$\beta$.

• Proceedings of the Korea Environmental Mutagen Society Conference
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• 2001.10b
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• pp.201-201
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• 2001

• Proceedings of the PSK Conference
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• 2001.10a
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• pp.173.1-173.1
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• 2001

• Proceedings of the PSK Conference
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• 2002.10a
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• pp.328.3-329
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• 2002

A convenient bioassay of nerve growth factor(NGF) is essential for assessing its potency during the course of product development and quality controls afterwards. We have set up a cell-based bioassay for determining the potency of recombinant NGF using rat pheochromocytoma (PC12) cells. Cell survival was measured by monitoring the reduction of the alamarBlue$^{TM}$ dye by living cells. (omitted)d)

• Korean Journal of Fisheries and Aquatic Sciences
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• v.34 no.6
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• pp.584-587
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• 2001

The effects of tributyltin (TBT), triphenyltin (TPhT) and Aroclor 1254 on germinal vesicle breakdown (GVBD) and ovulation of olive flounder (Paralichthys olivaceus) were investigated in in vitro bioassay. TBT, TPhT and Aroclor 1254 showed the inhibition effects on GVBD and ovulation in response to HCG. The oocyte response appeared to be more sensitive to TBT than Aroclor 1254. TBT was more effective in inhibition GVBD at concentrations of 0.1 and 1 ppm. However, no significant inhibition was obseued in concentrations tested ($0.0001\~1\;ppm$). Significant inhibition of ovulation in response to HCG occurred at TBT (0.01, 0.1, 1 ppm), TPhT (0.01, 0.1, 1 ppm) and Aroclor 1254 (0.01, 1 ppm, except 0,1 ppm), compared to HCG control, The lowest ovulation rate was measured at 1 ppm TBT, These data suggest that TBT (or TPhT) could possibly interfere the actions of progestogens to induce GVBD and ovulation in in vitro bioassay system.