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Recombinant human FIX concentrates (rhFIX) are essential in the treatment and prevention of bleeding in the bleeding disorder haemophilia B. However, due to the complex nature of FIX production yields are low which leads to high treatment costs. Here we report the production of rhFIX with substantially higher yield by co-expressing human FIX with GGCX (γ-glutamyl carboxylase), VKOR (vitamin K epoxide reductase) and furin (paired basic amino acid cleaving enzyme) in Chinese hamster ovary (CHO) cells. Our results show that controlled co-expression of GGCX with FIX is critical to obtain high rhFIX titre, and, that co-expression of VKOR further increased the yield of active rhFIX. Furin co-expression improved processing of the leader peptide of rhFIX but had a minor effect on yield of active rhFIX. The optimal expression level of GGCX was surprisingly low and required unusual engineering of expression vector elements. For VKOR and furin the control of expression was less critical and could be achieved by standard vector element. Using our expression vectors an rhFIX-producing clone with an expression level of up to 30 mg/L of active rhFIX was obtained. In addition an efficient single step purification method was developed to obtain pure and active rhFIX with up to 94 % yield.

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We present a systematic study of late Paleocene macrofauna from methane seep carbonates and associated driftwood in the shallow marine Basilika Formation, Spitsbergen, Svalbard. The fauna is composed of 22 taxa, comprising one brachiopod, 14 bivalves, three gastropods, three crustaceans, and one bony fish. The reported fish remains are among the first vertebrate body fossils from the Paleogene of Spitsbergen. One genus is new: the munidid decapod Valamunida Klompmaker and Robins gen. nov. Four new species are described: the terebratulide brachiopod Neoliothyrina nakremi Bitner sp. nov., the protobranch bivalve Yoldiella spitsbergensis Amano sp. nov., the xylophagain bivalve Xylophagella littlei Hryniewicz sp. nov., and the munidid decapod Valamunida haeggi Klompmaker and Robins gen. et sp. nov. New combinations are provided for the mytilid bivalve Inoperna plenicostata, the thyasirid bivalve Rhacothyas spitzbergensis, the ampullinid gastropod Globularia isfjordensis, and the munidid decapod Protomunida spitzbergica. Thirteen taxa are left in open nomenclature. The fauna contains a few last occurrences of Cretaceous survivors into the Paleocene, as well as first occurrences of Cenozoic taxa. It is composed of chemosymbiotic thyasirid bivalves and background species common in the northern Atlantic and Arctic during the Paleocene. Our results provide no evidence for a Paleocene origin of vesicomyid and bathymodiolin bivalves typical for Eocene and younger seep environments; instead, the Paleo cene seeps of the Basilika Formation are more similar to their Late Cretaceous equi valents rich in thyasirids.

A recently discovered Paleocene seep locality from Fossildalen on Spitsbergen, Svalbard, is described. This is one of a very few seep communities of the latest Cretaceous-earliest Palaeogene age, and the best preserved Paleocene seep community known so far. The seep