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    • 3. 发明授权
    • Cloned factor C cDNA of the Singapore horseshoe crab, Carcinoscorpius
rotundicauda and purification of factor C proenzyme
    • 新加坡马蹄蟹,Carcinoscorpius rotundicauda的克隆因子C cDNA和因子C酶原的纯化
    • US5716834A
    • 1998-02-10
    • US296014
    • 1994-08-19
    • Jeak Ling DingBow Ho
    • Jeak Ling DingBow Ho
    • C12N9/50C12N15/55C12Q1/37C12N15/63
    • C12N9/6408C12Y304/21084G01N33/579
    • Full-length and deletion subclones of cDNAs for Factor C of Carcinoscorpius rotundicauda are provided. These cDNAs have been cloned into .lambda.gt 22 and pGEM 11Zf(+). Further manipulations of the 5' and 3' ends of these cDNAs have been carried out, and these cDNAs have been further subcloned into other expression vectors such as pGEMEX-1, pET 3b, and the yeast shuttle vectors YEpsec 1 and pEMBLyex 4, and pPIC 9 and pHIL D2. Also provided are host cells transformed with expression vectors containing DNA molecules encoding proteins having Factor C-like enzymatic activity, methods of producing such proteins, methods for purifying Factor C zymogens, and methods for protecting Factor C zymogens from autoactivation by Gram negative bacterial endotoxin while the proenzyme is being purified and/or processed from amoebocyte lysates or from recombinant clones, or during storage or subsequent handling. This protection is afforded by the addition of 5-30% Me.sub.2 SO, which reversibly inhibits the Factor C zymogen.
    • 提供了番茄猕猴桃因子C的全长和缺失亚克隆。 这些cDNA已经克隆到λgt22和pGEM11Zf(+)中。 已经进行了这些cDNA的5'和3'端的进一步操作,并将这些cDNA进一步亚克隆到其他表达载体如pGEMEX-1,pET 3b和酵母穿梭载体YEpsec 1和pEMBLyex 4中,以及 pPIC 9和pHIL D2。 还提供了用含有编码具有因子C样酶活性的蛋白质的DNA分子转化的宿主细胞,产生这种蛋白质的方法,产生这种蛋白质的方法,纯化因子C酶原的方法,以及保护因子C酶原从革兰氏阴性细菌内毒素自动激活的方法, 正在从变形细胞裂解物或重组克隆纯化和/或加工,或在储存或后续处理过程中。 这种保护是通过加入5-30%的Me2SO,可逆地抑制因子C酶原来提供的。
    • 5. 发明申请
    • PATTERNED SOFT ADHESIVES AND METHOD OF MANUFACTURE
    • 图案软胶粘剂及其制造方法
    • US20100098846A1
    • 2010-04-22
    • US12568202
    • 2009-09-28
    • Jian Ling DingRengan KannabiranKathryn MoonRobert BraiewaBrian Thomas
    • Jian Ling DingRengan KannabiranKathryn MoonRobert BraiewaBrian Thomas
    • B05D5/10B05C1/08
    • C09J7/38C09J2201/28
    • A method is disclosed for making a soft adhesive tape. The method comprises providing a soft adhesive, rolling a gravure cylinder on the soft adhesive to meter the soft adhesive, depositing the metered soft adhesive from the gravure cylinder onto a carrier web, and transferring the soft adhesive to a backing substrate. In exemplary implementations, the method includes drying or curing the metered soft adhesive on the carrier web before transferring the soft adhesive to the backing substrate. A system is disclosed for making a layer of soft adhesive. The system comprises an applicator containing a soft adhesive, a gravure cylinder to meter the soft adhesive from the applicator, and a carrier web passing through the gravure cylinder, the carrier web supporting the metered soft adhesive deposited by the gravure cylinder. In exemplary implementations, a dryer may be included for curing the metered soft adhesive on the carrier web.
    • 公开了制造软胶带的方法。 该方法包括提供软粘合剂,在软粘合剂上滚动凹版滚筒以测量软粘合剂,将来自凹版滚筒的计量软粘合剂沉积到载体网上,并将软粘合剂转移到背衬基材上。 在示例性实施方案中,该方法包括在将软粘合剂转移到背衬基材之前干燥或固化载体网上的计量软粘合剂。 公开了一种用于制造软粘合剂层的系统。 该系统包括一个包含软粘合剂的涂抹器,一个用于对来自涂布器的柔软粘合剂进行计量的凹版滚筒,以及一个穿过凹版滚筒的载体网,支撑着由凹版滚筒沉积的计量软粘合剂的载体网。 在示例性实施方案中,可以包括干燥器以固化载体网上的计量软粘合剂。
    • 9. 发明授权
    • Assays for endotoxin
    • 内毒素检测
    • US06645724B1
    • 2003-11-11
    • US09287368
    • 1999-04-07
    • Jeak Ling DingBow Ho
    • Jeak Ling DingBow Ho
    • A61K3902
    • G01N33/579C07K14/43509C12N2799/026
    • The horseshoe crab, Carcinoscorpius rotundicauda Factor C cDNA (CrFC21) has been cloned into a shuttle baculoviral vector and another vector suitable for expression in insect cells. The recombinant baculoviral DNA was then transfected into the insect cells for expression of recombinant Factor C. Recombinant Factor C was found to be immunoreactive and is capable of binding both free and bound/immobilized lipid A. It is enzymatically active when triggered by LPS. The rFC is probably of the two-chain form, being cleaved into the heavy and light chains after activation by Gram negative bacterial endotoxin. As low as 0.01 pg (0.001 ng/ml) of LPS was detectable by the rFC, thus, indicating its potentials as a novel generation of “limulus amoebocyte lysate.”
    • 马蹄蟹Carcinoscorpius rotundicauda因子C cDNA(CrFC21)已经克隆到穿梭杆状病毒载体和另一种适合在昆虫细胞中表达的载体。 然后将重组杆状病毒DNA转染到昆虫细胞中用于表达重组因子C.重组因子C被发现是免疫反应性的,并且能够结合游离和结合/固定的脂质A.当LPS诱导时,其酶促活性。 rFC可能是双链形式,在革兰氏阴性细菌内毒素激活后被切割成重链和轻链。 通过rFC检测到低达0.01pg(0.001ng / ml)的LPS,因此,表明其作为新一代“鲎变形细胞裂解物”的潜力。
    • 10. 发明授权
    • Expression of Carcinoscorpius rotundicauda Factor C in eukaryotes
    • Carcinoscorpius rotundicauda因子C在真核生物中的表达
    • US5985590A
    • 1999-11-16
    • US877620
    • 1997-06-18
    • Jeak Ling DingBow Ho
    • Jeak Ling DingBow Ho
    • C12N9/50C12N15/55C12Q1/37
    • C12N9/6408C12Y304/21084G01N33/579
    • CrFC21 cDNA was cloned into two mammalian vectors: pCIneo and pCDNAI, both of which carry the strong CMV promoter for expression in mammalian cell lines. Various CrFC cDNA constructs transformed into P. pastoris and S. cerevisiae were expressed to yield full-length recombinant Factor C (rCrFC) protein of .about.130 kDa which is immunoreactive. The rCrFC is expressed in an intracellular, insoluble form. Intracellular localization of the nascent protein provides protection from premature digestion by proteases secreted by the host cell. Subsequent to its synthesis, rCrFC is solubilized and purified under pyrogen-free conditions. Using established protocols, the protein can be denatured and renatured to recover its biological functionality. By manipulation of the 5' end of CrFC26, truncated constructs containing this cDNA are expressed by S. cerevisiae to give immunoreactive rCrFC. The rCrFC produced from both CrFC21 and CrFC26 constructs, solubilized by Triton X-100 or SDS, is found to be immunoreactive. Solubilized rCrFC was purified as a proenzyme and reversibly protected from activation by addition of Me.sub.2 SO.
    • 将CrFC21 cDNA克隆到两个哺乳动物细胞系中的pCIneo和pCDNAI两个哺乳动物载体,两者都携带强CMV启动子进行表达。 转化到巴斯德毕赤氏酵母和酿酒酵母中的各种CrFC cDNA构建体被表达以产生具有免疫反应性的DIFFERENCE 130kDa的全长重组因子C(rCrFC)蛋白。 rCrFC以细胞内,不溶性形式表达。 新生蛋白质的细胞内定位提供了保护免受由宿主细胞分泌的蛋白酶的过早消化。 在合成之后,rCrFC在无热原条件下溶解和纯化。 使用既定的方案,蛋白质可以变性和复性以恢复其生物学功能。 通过操纵CrFC26的5'末端,含有该cDNA的截短构建体由酿酒酵母表达,得到免疫反应性rCrFC。 发现由Triton X-100或SDS溶解的CrFC21和CrFC26构建体产生的rCrFC是免疫反应性的。 将溶解的rCrFC纯化为酶原,并通过加入Me 2 SO可逆地保护免于活化。