• 제목/요약/키워드: Endogenous Growth

검색결과 287건 처리시간 0.024초

Plasminogen kringle 5 재조합 단백질에 의한 ERK1/2 활성화 및 세포골격 재배열 억제 (Inhibition of ERK1/2 Activation and Cytoskeleton Rearrangement by the Recombinant Protein of Plasminogen Kringle 5)

  • 하정민;김현경;김명래;조영애
    • 생명과학회지
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    • 제16권7호
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    • pp.1199-1206
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    • 2006
  • Plasminogen kringle 5는 plasminogen kringles 1-4로 구성된 내생의 혈관 신생 억제제인 angiostatin과 같이 내피세포의 분열을 강력하게 억제한다고 알려져 있다. 본 연구에서는 plasminogen kringle 5의 재조합 단백질을 효모 발현 체계에서 생산하여 내피세포의 이동에 대한 저해 효과와 이에 대한 작용기전을 조사하였다 재조합 단백질 PK5는 plasminogen의 Thr456에서 Phe546까지 이르는 cDNA 부분을 ${\alpha}-factor$ prepro-peptide의 분비 신호 서열 뒤에 도입하여 Pichia pastoris GS115에서 발현시켰다. 메탄올 유도 후 얻은 배양액을 S-spin column을 이용하여 정제하였다. 정제된 단백질을 SDS-PACE하였을 때 약 10kDa의 단일 밴드를 나타냄을 확인할 수 있었다. 정제된 PK5는 bFGF나 VEGF에 의해 유도된 인간의 제대 유래 내피 세포의 이동을 약 500nM의 $IC_{50}$ 값으로 농도 의존적으로 감소시켰다. 내피 세포에 PK5 500M을 처리한 결과 bFGF에 의해 유도된 ERK1/2의 인산화를 감소시켰다 또한, PK5는 bFGF에 의해 유도된 내피세포의 골격 재형성을 강력하게 억제하는 것으로 관찰되었다. 따라서, 이러한 결과들은 효모 생산 PK5가 내피세포의 이동을 효과적으로 억제하며, 이는 ERK1/2의 활성과 세포골격의 재배열을 억제함으로써 나타나는 것으로 부분적으로 설명될 수 있다.

체외배양 중인 생쥐 난소에서 초기난포 조절인자의 발현 (mRNA Expression of the Regulatory Factors for the Early Folliculogenesis in vitro)

  • 윤세진;김기령;정형민;윤태기;차광렬;이경아
    • Clinical and Experimental Reproductive Medicine
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    • 제32권3호
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    • pp.207-216
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    • 2005
  • Objective: To understand the crucial requirement for the normal early folliculogenesis, we evaluated molecular as well as physiological differences during in vitro ovarian culture. Among the important regulators for follicle development, anti-Müllerian hormone (AMH) and FSH Receptor (FSHR) have been known to be expressed in the cuboidal granulosa cells. Meanwhile, it is known that c-kit is germ cell-specific and GDF-9 is also oocyte-specific regulator. To evaluate the functional requirement for the competence of normal follicular development, we investigated the differential mRNA expression of several factors secreted from granulosa cells and oocytes between in vivo and in vitro developed ovaries. Materials and Methods: Ovaries from ICR neonates (the day of birth) were cultured for 4 days (for primordial to primary transition) or 8 days (for secondary follicle formation) in ${\alpha}$-MEM glutamax supplemented with 3 mg/ml BSA without serum or growth factors. The mRNA levels of the several factors were investigated by quantitative real-time PCR analysis. Freshly isolated 0-, 4-, and 8-day-old ovaries were used as control. Results: The mRNA of AMH and FSHR as granulosa cell factors was highly increased according to the ovarian development in both of 4- and 8-day-old control. However, the mRNA expression was not induced in both of 4- and 8-day in vitro cultured ovaries. The mRNA expression of GDF-9 known to regulate follicle growth as an oocyte factor was different between in vivo and in vitro developed ovaries. In addition, the transcript of GDF-9 was expressed in the primordial follicles of mouse ovaries. The mRNA expression of c-kit was not significantly different during the early folliculogenesis in vitro. Conclusion: This is the first report regarding endogenous AMH and FSHR expression during the early folliculogenesis in vitro. In conclusion, it will be very valuable to evaluate cuboidal granulosa cell factors as functional marker(s) for normal early folliculogenesis in vitro.

딸기 '설향' 육묘기 양분 공급 중단 시기가 자묘 생육 및 수량에 미치는 영향 (Effect of Timing of Nutrient Starvation during Transplant Production on the Growth of Runner Plants and Yield of Strawberry 'Seolhyang')

  • 김대영;채원병;곽정호;박수형;정승룡;최종명;윤무경
    • 생물환경조절학회지
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    • 제22권4호
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    • pp.421-426
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    • 2013
  • 국내에서 육성된 딸기 품종인 '설향'을 대상으로 비가림 포트 육묘 시 양분 공급 중단 시기에 따른 자묘의 생육과 정식 후 수량을 검토하였다. 자묘 받기를 완료한 후 양분을 EC $0.8dS{\cdot}m^{-1}$ 수준으로 공급하다가 7월 25일부터 9월 5일에 걸쳐 약 10일 간격으로 양분 공급을 각각 중단하고 이후 수분만 공급하였다. 그 결과 육묘기 양분 공급 중단 시기가 7월 25일로 빠른 경우에 엽면적등 지상부 생육은 억제되었으나 근중은 증가하여 T/R율이 크게 감소하였고 엽록소 함량이 낮아지는 경향을 보였다. 또한 화아 분화가 촉진되어 정식 후의 출뢰, 개화 및 수확기가 앞당겨졌으나 8월 중순 이후까지 양분 공급이 지속될 경우에 수확 시기가 2주 이상 지연되었다. 이에 따라 7월 25일에 양분 공급을 중단한 처리구에서 1월 하순 및 2월 하순까지의 1주당 누적 상품과 수량은 각각 169 및 266g으로 나타났으며 9월 5일 처리구와 비교하여 각각 71 및 12% 수준의 유의한 수량 증가를 보였다. 따라서 육묘 후기 자묘의 적절한 양분 공급 중단으로 체내 질소 수준을 효과적으로 감소시킴으로써 딸기의 화아 분화를 촉진하고 수확 시기를 앞당겨 딸기 가격이 높게 형성되는 동절기 수량 및 소득을 증대시킬 수 있을 것으로 기대된다.

Imaging Neuroreceptors in the Living Human Brain

  • Wagner Jr Henry N.;Dannals Robert F.;Frost J. James;Wong Dean F.;Ravert Hayden T.;Wilson Alan A.;Links Jonathan M.;Burns H. Donald;Kuhar Michael J.;Snyder Solomon H.
    • 대한핵의학회지
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    • 제18권2호
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    • pp.17-23
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    • 1984
  • For nearly a century it has been known that chemical activity accompanies mental activity, but only recently has it been possible to begin to examine its exact nature. Positron-emitting radioactive tracers have made it possible to study the chemistry of the human mind in health and disease, using chiefly cyclotron-produced radionuclides, carbon-11, fluorine-18 and oxygen-15. It is now well established that measurable increases in regional cerebral blood flow, glucose and oxygen metabolism accompany the mental functions of perception, cognition, emotion and motion. On May 25, 1983 the first imaging of a neuroreceptor in the human brain was accomplished with carbon-11 methyl spiperone, a ligand that binds preferentially to dopamine-2 receptors, 80% of which are located in the caudate nucleus and putamen. Quantitative imaging of serotonin-2, opiate, benzodiazapine and muscarinic cholinergic receptors has subsequently been accomplished. In studies of normal men and women, it has been found that dopamine and serotonin receptor activity decreases dramatically with age, such a decrease being more pronounced in men than in women and greater in the case of dopamine receptors than serotonin-2 receptors. Preliminary studies in patients with neuropsychiatric disorders suggests that dopamine-2 receptor activity is diminished in the caudate nucleus of patients with Huntington's disease. Positron tomography permits quantitative assay of picomolar quantities of neuro-receptors within the living human brain. Studies of patients with Parkinson's disease, Alzheimer's disease, depression, anxiety, schizophrenia, acute and chronic pain states and drug addiction are now in progress. The growth of any scientific field is based on a paradigm or set of ideas that the community of scientists accepts. The unifying principle of nuclear medicine is the tracer principle applied to the study of human disease. Nineteen hundred and sixty-three was a landmark year in which technetium-99m and the Anger camera combined to move the field from its latent stage into a second stage characterized by exponential growth within the framework of the paradigm. The third stage, characterized by gradually declining growth, began in 1973. Faced with competing advances, such as computed tomography and ultrasonography, proponents and participants in the field of nuclear medicine began to search for greener pastures or to pursue narrow sub-specialties. Research became characterized by refinements of existing techniques. In 1983 nuclear medicine experienced what could be a profound change. A new paradigm was born when it was demonstrated that, despite their extremely low chemical concentrations, in the picomolar range, it was possible to image and quantify the distribution of receptors in the human body. Thus, nuclear medicine was able to move beyond physiology into biochemistry and pharmacology. Fundamental to the science of pharmacology is the concept that many drugs and endogenous substances, such as neurotransmitters, react with specific macromolecules that mediate their pharmacologic actions. Such receptors are usually identified in the study of excised tissues, cells or cell membranes, or in autoradiographic studies in animals. The first imaging and quantification of a neuroreceptor in a living human being was performed on May 25, 1983 and reported in the September 23, 1983 issue of SCIENCE. The study involved the development and use of carbon-11 N-methyl spiperone (NMSP), a drug with a high affinity for dopamine receptors. Since then, studies of dopamine and serotonin receptors have been carried out in over 100 normal persons or patients with various neuropsychiatric disorders. Exactly one year later, the first imaging of opitate receptors in a living human being was performed [1].

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Effects of Single Cell Protein Replacing Fish Meal in Diet on Growth Performance, Nutrient Digestibility and Intestinal Morphology in Weaned Pigs

  • Zhang, H.Y.;Piao, X.S.;Li, P.;Yi, J.Q.;Zhang, Q.;Li, Q.Y.;Liu, J.D.;Wang, G.Q.
    • Asian-Australasian Journal of Animal Sciences
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    • 제26권9호
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    • pp.1320-1328
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    • 2013
  • Three experiments were conducted to evaluate the ME value, standardized ileal digestibility (SID) of amino acids (AA) of fish meal, and the effects of single cell protein (Prosin and Protide) replacing fish meal in diet on growth performance, nutrient digestibility and intestinal morphology in weaned piglets. In Exp. 1, twenty-four barrows with initial BW of $30.8{\times}2.6kg$ were allotted to one of four dietary treatments. Diet 1 contained corn as the only energy source. The other three diets replaced 20% of the corn in diet 1 with one of the three protein feeds (fish meal, Prosin and Protide), and the DE and ME contents were determined by difference. In Exp. 2, eight barrows (initial BW of $25.6{\pm}3.2kg$) were fitted with ileal T-cannulas and allotted to a replicated $4{\times}4$ Latin square design. Three cornstarch-based diets were formulated using each of the protein feeds as the sole source of AA. A nitrogen-free diet was also formulated to measure endogenous losses of AA. In Exp. 3, one hundred and eighty piglets (initial BW of $7.95{\pm}1.59kg$) weaned at $28{\times}2d$ were blocked by weight and assigned to one of five treatments for a 28-d growth performance study, each treatment was fed to six pens with six pigs (three barrows and three gilts) per pen. The five treatments consisted of the control group (CON), which was a corn-soybean meal diet containing 5% fish meal, and the other four treatments, which replaced a set amount of fish meal with either Prosin (2.5% or 5%) or Protide (2.5% or 5%). The diets were formulated to provide same nutrient levels. The results showed that on a DM basis, both of the DE and ME contents were lower in Prosin and Protide than that of fish meal (p<0.05). The SID of CP and all essential AA were greater in fish meal than in Prosin and Protide (p<0.05). The pigs fed CON diet had greater weight gain and lower feed conversion rate (FCR) than pigs fed 5% Prosin and 5% Protide diets (p<0.05). The digestibility of CP was greater in pigs fed CON, 2.5% Prosin and 2.5% Protide diets than the pigs fed 5% Prosin and 5% Protide diets (p<0.05). Villus height in jejunum and ileum, and villus height to crypt depth ratio in the jejunum were higher (p<0.05) in pigs fed CON, 2.5% Prosin and 2.5% Protide diets compared with the 5% Prosin and 5% Protide diets. Pigs fed CON diet had greater villus height to crypt depth ratio in the ileum than the pigs fed 5% Prosin and 5% Protide diets (p<0.05). In conclusion, although Prosin and Protide contained lower ME content and SID of AA than fish meal, Prosin and Protide replacing 50% of fish meal in diet with identical nutrient levels could obtain similar performance, nutrient digestibility and intestinal morphology in weaned pigs.

WASP7 모형을 이용한 임하호 수질모의에 관한 연구 (The Research about the Water Quality Prediction at Imha Reservoir Using a WASP7 Model)

  • 안승섭;서명준;정도준;박노삼
    • 한국환경과학회지
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    • 제17권6호
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    • pp.611-621
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    • 2008
  • This study intends to provide the necessary basic data needed for predicting the water quality and examining changes in water quality on the basis of the hydrological changes: an outflow or the character of a flow by investigating the interaction of the parameters through the estimation of optimal parameters need for predicting the water quality of the dam basin and the sensitivity among those estimated parameters. Im-Ha Dam in the upstream area of the Nakdong River was selected for analysis, and the water quality survey data necessary for parameter estimation was based on the monthly water quality data (water temperature, BOD, T-N and T-P) between December 1, $2005{\sim}$November 31, 2006. K1C(the saturated growth rate of plant plankton), K1RC (endogenous respiratory quotient of plankton), KDC(deoxidized ratio), K71C(minealized ratio of dissolved organic phosphorus), K83C(mineralized ratio of dissolved organic nitrogen) have been considered as the factors of the water quality performed in this water quality simulation, that is, the most effective parameters on BOD, T-N and T-P. In the result of the analysis of the sensitivity, KDC(deoxidized ratio) was the most sensitively reacted parameter on BOD and it was K71C(mineralized ratio of dissolved organic phosphorus) and K83C(mineralized ratio of dissolved organic nitrogen) on T-N and T-P. It is considered that it will be possible to apply the most optimal parameter to an analysis of the water quality simulation at Im-Ha Ho basin in the goal year by examining the interaction of the parameters through the parameters sampling which are able to applicable to prediction of the water quality and the analysis of the its sensitivity, in the future, also the analysis on the basis of the hydrological conditions: an outflow or the character of a flow will be needed.

Knock-in of Enhanced Green Fluorescent Protein or/and Human Fibroblast Growth Factor 2 Gene into β-Casein Gene Locus in the Porcine Fibroblasts to Produce Therapeutic Protein

  • Lee, Sang Mi;Kim, Ji Woo;Jeong, Young-Hee;Kim, Se Eun;Kim, Yeong Ji;Moon, Seung Ju;Lee, Ji-Hye;Kim, Keun-Jung;Kim, Min-Kyu;Kang, Man-Jong
    • Asian-Australasian Journal of Animal Sciences
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    • 제27권11호
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    • pp.1644-1651
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    • 2014
  • Transgenic animals have become important tools for the production of therapeutic proteins in the domestic animal. Production efficiencies of transgenic animals by conventional methods as microinjection and retrovirus vector methods are low, and the foreign gene expression levels are also low because of their random integration in the host genome. In this study, we investigated the homologous recombination on the porcine ${\beta}$-casein gene locus using a knock-in vector for the ${\beta}$-casein gene locus. We developed the knock-in vector on the porcine ${\beta}$-casein gene locus and isolated knock-in fibroblast for nuclear transfer. The knock-in vector consisted of the neomycin resistance gene (neo) as a positive selectable marker gene, diphtheria toxin-A gene as negative selection marker, and 5' arm and 3' arm from the porcine ${\beta}$-casein gene. The secretion of enhanced green fluorescent protein (EGFP) was more easily detected in the cell culture media than it was by western blot analysis of cell extract of the HC11 mouse mammary epithelial cells transfected with EGFP knock-in vector. These results indicated that a knock-in system using ${\beta}$-casein gene induced high expression of transgene by the gene regulatory sequence of endogenous ${\beta}$-casein gene. These fibroblasts may be used to produce transgenic pigs for the production of therapeutic proteins via the mammary glands.

Expression of Egr3 in mouse gonads and its localization and function in oocytes

  • Shin, Hyejin;Seol, Dong-Won;Nam, Minyeong;Song, Haengseok;Lee, Dong Ryul;Lim, Hyunjung Jade
    • Asian-Australasian Journal of Animal Sciences
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    • 제30권6호
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    • pp.781-787
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    • 2017
  • Objective: The early growth response (Egr) family consists of four members (Egr1, Egr2, Egr3, and Egr4) that are zinc finger transcription factors. Among them, Egr3 is involved in transcriptional regulation of target genes during muscle spindle formation and neurite outgrowth. We previously showed that the immunoreactive Egr3 is localized on oocyte spindle and accumulate near the microtubule organizing center during meiosis I in mice. Egr3 was also shown to be localized on spermatocytes. We herein investigated if Egr3 is expressed in mouse gonads and if Egr3 blockade results in any defect in oocyte maturation. Methods: Expression of Egr3 in mouse gonads was examined by reverse transcription-polymerase chain reaction. Full-length Egr3 and truncated Egr3 (${\Delta}Egr3$) complementary RNAs (cRNAs) with Xpress tag at N-terminus and DsRed2 at C-terminus, and small interfering RNA (siRNA) targeting Egr3 were microinjected into mouse oocytes at germinal vesicle stage. Localization of microinjected Egr3 was examined by confocal live imaging and immunofluorescence staining. Results: Egr3 mRNA was detected in mouse ovaries and testes from 1 to 4 week-old mice. An uncharacterized longer transcript containing 5'untranslated region was also detected in 3 and 4 week-old gonads. Microinjected Xpress-Egr3-DsRed2 or Xpress-${\Delta}Egr3$-DsRed2 localized to nuclei and chromosomes during meiotic progression. Microinjection of these cRNAs or Egr3 siRNA in oocytes did not affect meiotic maturation. Immunofluorescence staining of Egr3 in Xpress-${\Delta}Egr3$-DsRed2-injected oocytes showed a positive signal only on meiotic spindle, suggesting that this antibody does not detect endogenous or exogenous Egr3 in mouse oocytes. Conclusion: The results show that Egr3 localizes to chromosomes during meiotic progression and that certain antibodies may not faithfully represent localization of target proteins in oocytes. Egr3 seems to be dispensable during oocyte maturation in mice.

전략적 지역혁신체재 구축을 위한 지역혁신협의회 조직에 관한 연구 - 강원도 사례를 중심으로 (A Study on the Organization Design of Regional Innovation Councils for Establishing Strategic Regional Innovation Systems: Based on Kang-Won Province Case)

  • 이주헌
    • 산학경영연구
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    • 제21권2호
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    • pp.195-214
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    • 2008
  • 지난 2004년 이후로 정부는 지역의 내생적 자립과 발전을 위한 핵심역량과 성장동력을 창출하기 위해 지역혁신체계를 구축하기 위한 정책을 추진하였고 추진주체로서 산학연 전문가를 중심으로 구성된 지역혁신협의회를 만들었다. 당시 강원도의 지역혁신협의회 조직은 운영위원회와 사무국을 중심으로 기획조정분과, 전략산업분과, 문화관광산업분과, 지연산업분과, 지역인적자원개발분과 등으로 구성되어 지역혁신정책을 수립, 검토, 심의, 평가, 확정하는 업무를 담당하는 등 의욕적으로 추진되었다. 하지만, 기존 정부조직과의 역할차별화 문제, 균형발전위원회의 예산확보문제, 전략적 조직체계 문제, 의사결정구조 문제등으로 애초에 기획했던 지역혁신협의회의 기능과 역할과는 달리 미래지향적 전략설정과 혁신적 지역산업정책의 추진은 어렵게 되고 지역에서 기존에 추진하고 있던 산업과 연구자원에 근거하여 전략산업이 선정되고 예산배분이 결정되었다. 성공적이지 못한 결과에도 불구하고 오늘날과 같은 혁신사회에서는 지역의 혁신정책을 결정하고 심의, 조정할 수 있는 지역혁신협의회와 같은 조직의 역할과 필요성이 점점 중요해지고 있는 추세이다. 아마 차기정부에서도 비슷한 역할을 담당할 혁신과 창업의 추진체가 앞으로도 계속 출현할 것으로 예상한다. 본 논문의 목적은 지역혁신협의회의 조직구조를 분석하고 기존 조직의 전략과 조직구조에 대한 문제점을 도출하고 이에 대한 해결책을 제시함으로써 바람직한 혁신추진체의 역할과 모습을 재정립하는 것이다. 본 논문에서는 저자가 속한 강원도 지역혁신협의회의 조직구조와 분과의 역할을 체계적으로 분석하고 문제점을 식별, 논의하고 바람직한 추진체가 갖추어야 할 사항과 시사점을 제시하고자 하였다.

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An engineered PD-1-based and MMP-2/9-oriented fusion protein exerts potent antitumor effects against melanoma

  • Wei, Mulan;Liu, Xujie;Cao, Chunyu;Yang, Jianlin;Lv, Yafeng;Huang, Jiaojiao;Wang, Yanlin;Qin, Ye
    • BMB Reports
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    • 제51권11호
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    • pp.572-577
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    • 2018
  • Recent studies showed that the PD-1/PD-L1 checkpoint blockade is a dramatic therapy for melanoma by enhancing antitumor immune activity. Currently, major strategies for the PD-1/PD-L1 blockade have mainly focused on the use of antibodies and compounds. Seeking an alternative approach, others employ endogenous proteins as blocking agents. The extracellular domain of PD-1 (ePD1) includes the binding site with PD-L1. Accordingly, we constructed a PD-1-based recombinantly tailored fusion protein (dFv-ePD1) that consists of bivalent variable fragments (dFv) of an MMP-2/9-targeted antibody and ePD1. The melanoma-binding intensity and antitumor activity were also investigated. We found the intense and selective binding capability of the protein dFv-ePD1 to human melanoma specimens was confirmed by a tissue microarray. In addition, dFv-ePD1 significantly suppressed the migration and invasion of mouse melanoma B16-F1 cells, and displayed cytotoxicity to cancer cells in vitro. Notably, dFv-ePD1 significantly inhibited the growth of mouse melanoma B16-F1 tumor cells in mice and in vivo fluorescence imaging showed that dFv-ePD was gradually accumulated into the B16-F1 tumor. Also the B16-F1 tumor fluorescence intensity at the tumor site was stronger than that of dFv. This study indicates that the recombinant protein dFv-ePD1 has an intensive melanoma-binding capability and exerts potent therapeutic efficacy against melanoma. The novel format of the PD-L1-blocked agent may play an active role in antitumor immunotherapy.