• BMB Reports
• /
• 제42권11호
• /
• pp.731-736
• /
• 2009

The generation of functional recombinant antibodies from hybridomas is necessary for antibody engineering. However, this is not easily accomplished due to high levels of aberrant heavy and light chain mRNAs, which require a highly selective technology that has proven complicated and difficult to operate. Herein, we attempt to use an alkaline phosphate (AP)-fused form of single-chain variable fragment (scFv) for the simple identification of a hybridoma-derived, functional recombinant antibody. As a representative example, we cloned the scFv gene from a hybridoma-producing mouse IgG against branched-chain keto acid dehydrogenase complex-E2 (BCKD-E2) into an expression vector containing an in-frame phoA gene. Functional recombinant antibodies were easily identified by conventional enzyme-linked immunosorbent assay (ELISA) by employing scFv-AP fusion protein, which also readily serves as a valuable immuno-detective reagent.

• 미생물학회지
• /
• 제23권3호
• /
• pp.172-176
• /
• 1985

본 연구는 효모세포 (Saccharomyces uvarum)의 ACPase, ALPase의 Isoenzyme type을 규명함과 아울러, iso-enzyme type중 어느 것이 constituitive 또는 repressible enzyme type인가를 조사하고, 각각의 isoenzyme활성도 비율을 살펴보아 배양조건(catabolic repression and derepression) 및 무기인산 제한에 따른 각 type의 세포내 조절기능을 분석하고자 하였다. Acid phosphatase는 또 다른 isoenzyme type이 발견되지 않았으나, alkaline phosphatase의 경우는 3가지 type이 p-NPP에 specificity를 지녔다. 세포질의 수용성단백질 분석결과 exponential phase의 세포는 주로 음전하를 띤 단백질이 높은 함량을 나타내었다. 당과 인산이 결핍된 배지에서 생육권(catabolic repression)세포에서 AL P Pase isoenzyme 중 type “B"의 활성도가 매우 높아졌으나, 완전배지에서 배양된 (catabolic derepression)세포에서는 type “B"의 활성도 감소 및 type “C"의 활성도 증가 현상을 볼 수 있었다. ALPase 중 “A" peak는 constituitive enzyme, “B" peak는 repressible enzyme, peak “C"는 L-histidinol phosphatase로 추정되었다.

• 약학회지
• /
• 제42권4호
• /
• pp.370-381
• /
• 1998

The effect of herbal medicine on osteoporosis was studied using ovariectomized rats as an animal model of Type I osteoporosis and SAM P6 mice as that of Type II. Each traditiona l boiling water extract of Achyranthis Radix, Psoraleae Radix, Rehmanniae Radix Preparat, Corni Fructus and Mycelia of Ganoderma, and systemic water fraction of Astragali Radix was given 5g(dried herbal weight)/kg/day p.o. for 30 days in each group of ovariectomized rats, SAM R1 and SAM P6. The extract of Cervi parvum Cornu was given for 14 days only. One ml of blood was taken by tail vein at day 0, 7, 14, 21 and 30 days after administration of the extract. Plasma levels of alkaline phosphatase, calcium, creatinine,inorganic phosphate, blood urea nitrogen, cortisol, total $T_3\;and\;total\;T_4$ were measured. In ovariectomized rats, administration of Achyranthis Radix or Corni Fructus decreased in alkaline phosphatase and that of Achyranthis Radix or Psoraleae Radix decreased in calcium comparing to the control (p<0.05). The administration of Psoralese Radix decreased in calcium and increased in urea comparing to day o(P<0.05)(Table I). There were not much changes in plasma calcium, inorganic phosphate, and alkaline phosphatase concentrations after uptake of these herbal medicine used in SAM P6(Table III). However, administration of Astragali Radix altered plasma inorganic phosphate and creatinine levels in SAM R1(p<0.01)(Table UU). The administration of Corni Fructus or Psoralease Radix induced the changes in plasma concentrations of cortisol, total $T_3$ and total $T_4$ in Type I(p<0.05) (Table IV). The uptake of Cervi parvum Cornu increased in total $T_3$ concentration and that of Mycelia of Ganodtragali Radix in SAM P6. However, the uptake of Mycelia of Ganoderma induced changes in cortisol and $T_4$ concentrations in SAM R1(p<0.05). Thus, there were significant differences in responses of herbal medicine in different types of osteoporosis.

• Journal of Nutrition and Health
• /
• 제28권9호
• /
• pp.817-824
• /
• 1995

This study was performed to evaluate the effect of dietary protein level on Ca efficiency in bone mineral density in growing male rats. Twenty male rate were divided into two groups. The rats in one group were fed on casein 20% diet as control group and the others were fed on casein 40% diet as protein group. All rats were fed on experimental diet and deionized water ad libitum for 9 weeks. The total body, spine and femur bone mineral density and bone mineral content were measured using dual energy-x ray absorptiometry. Urinary calcium, phosphate, pyridinoline and creatinine, serum calcium, phosphate, total protein, albumin, alkaline phosphatase(ALP) and osteocalcin were measured. Urinary Ca excretion, pyridinoline and crosslinks value and serum ALP content seem to be increased in high protein group. It appears that the growing rats in high protein group had a higher bone resprption and bone formation than those in control group. Animal fad a high protein diet had a siginficantly higher Ca efficiency in BMD, BMC of total body, spine and femur. The results of this show that increasing of dietary protein level (40%) is beneficial of improvement of Ca efficiency during growing period.

• Bulletin of the Korean Chemical Society
• /
• 제28권12호
• /
• pp.2315-2318
• /
• 2007

Epinephrine was determined using a lab-made chemiluminescence (CL) system with air pump. Luminolsodium IO4? chemiluminescence system was employed to produce the luminescence of epinephrine. In the reaction, epinephrine was oxidized to produce superoxide or singlet oxygen by periodate in alkaline solution, which enhanced CL of luminol. For optimization, various buffers, such as phosphate, borate, and tris, were studied in this experiment. Compared to NaOH, the phosphate and borate buffer showed better reproducibility with similar sensitivity. Small amount of sample, 22 μL, was required for a measurement. The limit of quantification for epinephrine was obtained to be ~10?9 g/mL after optimization.

• Bulletin of the Korean Chemical Society
• /
• 제29권5호
• /
• pp.943-947
• /
• 2008

A phosphate-specific and fluorescent probe was prepared for label-free phosphatase assays based on fluorescence polarization. By using the probe, dephosphorylation reactions of DNA and protein substrates by calf intestinal alkaline phosphatase (CIP) could effectively be monitored in real-time. Since this assay method does not require additional materials such as labeled substrates and phosphospecific antibodies to obtain fluorescence polarization signals, it is simple, cost-effective, and expected to be useful not only for measuring activity of phosphatases but also for high-throughput screening of phosphatase inhibitors.

• 미생물학회지
• /
• 제28권3호
• /
• pp.249-257
• /
• 1990

In the course of operating the accommodative and detoxifying mechanism against cadmium, its physio-biochemical changes were observed in Klebsiella aerogenes ATCC 10031. Changes of enzyme activity concerned phosphate metabolism, changes of phospholipid composition and in view of energy metabolism the changes of the nucleotide pool were examined. Activities of both alkaline and acid phosphatase were derepressed 4-10 folds under cadmium added cultures. Moreover, production of phospholipid such as lysophosphatidyl choline (LPC), phosphatidyl serine (PS) and phosphatidyl ethanolamone (PE) was increased and uridylate nucleotide pool was increased under Cd-surplus culture. These results i.e. overproduction of phosphatase catalyzing phosphate residue, increase of the production of PE and PS which have a close affinity with cadmium, and indrease of uridylate nucleotide pool used as a carrier of polysaccharide synthesis like bacterial capsule exhibited cellular responses for active defence against Cd-pressure. It was postulated that these phenomena should play another assistant roles in Cd-detoxifing mechanism.

• Journal of Industrial and Engineering Chemistry
• /
• 제68권
• /
• pp.220-228
• /
• 2018

In this study, we found a simple surface biofunctionalization of biphasic calcium phosphate (BCP) based on the high affinity between alendronate and the calcium ions of BCP, and the strong interaction between heparin and bone morphogenic protein-2 (BMP-2). The biofunctionalized BCP did not be precipitated well and display a remarkable enhancement of osteogenic activity of human adipose-derived stem cells by showing increased alkaline phosphatase (ALP), calcium deposition and osteogenic-related genes (i.e., Runx-2, ALP, osteocalcin, and osteopontin), and bone regeneration in the calvarial defect model. Therefore, this simple surface technique can be used to easily functionalize various calcium phosphates.

• 한국환경농학회지
• /
• 제30권3호
• /
• pp.261-267
• /
• 2011

인산에 의한 카드뮴의 식물저감 효과를 증진시키기 위해 서로 다른 특성의 두 인산제재인 용과린과 $K_2HPO_4$를 선정하여 축분퇴비와 혼용하여 처리한 후 알타리무 내 카드뮴 흡수특성과 토양 내 카드뮴의 특성을 조사하였다. $K_2HPO_4$는 토양의 pH와 음하전도를 증대시켜 토양 내 유효태 카드뮴 함량과 알타리무내 카드뮴 흡수농도를 저감시키는 효과를 나타내었다. 반면 용과린은 토양의 pH와 음하전도를 감소시켜 유효태 카드뮴 함량과 알타리무 내 카드뮴 함량을 증가시키는 효과를 나타내었다. 축분퇴비와 인산비료와의 혼용은 인산 제재의 특성에 관계없이 인산비료를 단독으로 시용하였을 때 보다 알타리무 내 카드뮴 흡수농도와 토양 내 유효태 카드뮴 함량을 감소시키는 효과가 훨씬 우수한 것으로 나타났다. 인산비료나 축분퇴비의 시용에 의해 토양 내 인산과 카드뮴의 침전반응에 의한 난용성의 카드뮴 화합물의 형성은 나타나지 않았다. 토양의 pH와 음하전도는 토양 내 유효태 카드뮴의 함량과 알타리무 내 카드뮴의 함량과 고도로 유의한 부의 상관관계를 나타내었다. 본 연구의 결과를 통하여 인산비료와 축분퇴비에 의한 카드뮴의 부동화는 인산과 카드뮴의 직접적인 침전반응에 의한 것이라기보다는 토양의 pH와 음하전도의 증대에 기인된 카드뮴의 흡착증대에 의한 것으로 판단되어진다.

• Asian-Australasian Journal of Animal Sciences
• /
• 제32권4호
• /
• pp.527-532
• /
• 2019

Objective: Phosphorous (P) sources with greater bioavailability might increase animal production efficiency and decrease environmental pollution. The objective of current study was to determine animal performance, nutrient digestibility, blood metabolites and fecal P concentration in finishing lambs fed a diet with either di-calcium phosphate (DCP) or di-ammonium phosphate (DAP) as a P source. Methods: Twelve 4-month-old male lambs (initial body weight $24.87{\pm}3.4kg$) were randomly allocated to a diet with either DCP or DAP (~261 g/kg of total diet P) fed ad libitum for 93 days. Diets were iso-nitrogenous and iso-energetic and had same calcium (Ca) and P concentrations. Results: The DAP contained 19.7 g/kg of dry matter (DM) Ca, 185.4 g/kg DM P and 14,623 ppm fluorine, while DCP contained 230.3 g/kg DM Ca, 195.2 g/kg DM P and 1,039 ppm fluorine. The diet with DAP contained 60 ppm fluorine while the diet with DCP contained 13 ppm fluorine. Lambs fed the diet with DAP tended to have a greater daily DM intake compared to those fed diet with DCP (p = 0.09). Lambs fed DAP had greater plasma P concentration and alkaline phosphatase activity ($p{\leq}0.01$) compared with lambs fed DCP. Dry matter and organic matter digestibility of the diets were similar between two treatments at days 60 and 90, while they were greater in lambs fed DCP (p<0.05) at day 30 of the trial. Feeding DAP increased P digestibility (58.7% vs 50.2%; p<0.05) and decreased fecal P concentration in lambs compared with feeding DCP (3.1 vs 3.8 g/kg DM; p<0.05). Conclusion: Providing ~261 g/kg of total diet P as DAP in the diet of finishing lambs improved the bioavailability of P in the body and decreased excretion of P in feces without affecting lamb performance.