제조기술 번역에 대해서 알아 보겠습니다(한영번역)
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제조기술 번역(한국어 원본)Polymer를 이용한 Multi-layer film 제조 기술은 그동안 수많은 연구가 진행되어 왔다. 그러나 polymer 대신 bioactive material을 patterned multi-layer로 만드는데 사용하면 drug delivery vehicle, biosensor, nanoelectronic devices등의 의료분야에 더 많이 이용될 수 있을 것이다. 현재까지 Bioactive material을 patterned multi-layer로 만든 연구는 모두 protein particle 생산 후 추가적인 modification공정이 들어가고, patterning의 높이 (layer개수)가 낮거나 patterning control이 매우 힘들었다. 그러나 우리가 사용한 pH induced charge-charge repulsion and interaction을 이용하여 patterning을 하는 biocompatible multi-layer patterning process는 위의 문제점들을 모두 한번에 해결해 주며 추가적으로 protein particle을 probe로 사용하여 3D platform에 binding시켜 sensor로 활용한다는 점에서 Park et al. 이 언급한 세가지 장점(convenient and efficient control of antigen orientation upon the surface immobilization of antibodies, substantial increase in density and ratio of antigens to protein markers, 3-dimensional surface) 또한 갖는다. 또한 다양한 praticle을 사용하여 multi-detection 이 가능하게 할 수 있으며, non-specific binding을 막기위한 blocking과정이 필요없다는 장점이 있다. 그림과 같이 Ni palte 위에 일정한 간격으로 일정한 크기의gold dot을 만들어 patterned multi-layer를 위한 기초로 사용하였다. 그림1과 같이Gold dot에는 adenine의 gold에 대한 affinity를 이용하여 polyA를 binding시키고, 산화된 Ni위에는 Proteasome alpha subunit N-term의 His-tag을 이용하여 Proteasome을 binding 시킨다. 그 다음에 pH5 PBS로 buffer change한 recombinant DPS (DPS::BRCAA1)를 incubation하면 pH5에서는 그림과 같이 Proteasome alpha subunit과 DPS가 모두 강한 positive charge를 띄므로 DPS가 선택적으로 negative charge를 띄는polyA의 phosphate back bone에 binding하게 된다. Washing과정을 거치고 DNA double strand (DNA double strand of DPS in pH8 PBS)를 incubation하면 pH8에서는 Proteasome alpha subunit이 negative charge를 띄며, DPS 역시 weak negative charge를 띄지만 pH8에서도 DPS와 DNA는 binding을 매우 잘 한다는 연구결과도 있고 (우리 실험 결과도 역시 DPS와 DNA가 pH5, pH8에서 모두 binding을 매우 잘 함을 보여준다.) 상대적으로 proteasome alpha의 negative charge가 더 강하므로, DNA가 선택적으로 DPS와 binding을 한다. pH8에서는 아마도 DPS의 전체 net charge영향 보다는 N-term에 집중되어 있는 Lys의 국지적으로 강한 positive charge의 영향이 우세하여DPS와 DNA가 binding하는 것 같다. 이러한 과정을 반복하면 DPS와 DNA를 gold dot위에 선택적으로 쌓아 올려 patterned multi-layer of protein particle을 만들 수 있다. |
제조기술 번역(영어 번역본)Previously, there have been numerous studies on the multi-layer film manufacture technology using polymers. However, if a bioactive material is used to make a patterned multi-layer instead of a polymer, the patterned multi-layer can be used in more fields such as drug delivery vehicle, biosensor and nanoelectronic device. Up to now, studies on making a patterned multi-layer with bioactive materials all had difficulties due to the requirement of additional modification process after the manufacturing of protein particles and the extremely low patterning height (number of layers) or the difficulties in patterning control. However, the biocompatible multi-layer patterning process using the pH induced charge-charge repulsion and interaction used in this study solves all of the above problems. Also, this provides the three merits mentioned by Park et al. (convenient and efficient control of antigen orientation upon the surface immobilization of antibodies, substantial increase in density and ratio of antigens to protein markers, 3-dimensional surface) because it uses a protein particle as a probe to bind on the 3D platform and uses it as a sensor. Also, this study provides a way for multi-detection using various particles and also has benefits in a point that it does not require a blocking process for non-specific binding. Gold dots of a definite size are made on a Ni plate at definite intervals and the gold dots were used as the basis for the patterned multi-layer. As seen in Fig. 1, polyA was bound on the gold dot using the adenine’s affinity to gold and then the proteasome was bound on Ni using the His-tag of the proteasome alpha subunit N-term. Then the recombinant DPS (DPS::BRCAA1), which was buffered with pH5 PBS, was incubated. And because both the proteasome alpha subunit and DPS show strong positive charges in pH5, the DPS binds to the phosphate back bone of polyA, which is negatively charged selectively. When the DNA double strand (DNA double strand of DPS in pH8 PBS) is incubated after the washing process, the proteasome alpha subunit is negatively charged in pH8, and although DPS also is negatively charged at a very minimal level, there are studies that show DPS and DNA also bind very well in pH8 (this study also shows that DPS and DNA bind very well in pH5 and pH8), and when the negative charge of the proteasome alpha is relatively stronger, DNA selectively binds with DPS. The authors assume that the DPS and DNA bind together because the locally strong positive charge of Lys concentrated in the N-term is more effective than the effects of the net charge of DPS in pH8. Repeating this process, DPS and DNA can be selectively stacked on the gold dot to make a patterned multi-layer of protein particles. |
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