Recombinant Expression of Complex Proteins in Human Cell Lines.

Coagulation factors, as factor VII, VIII, and IX, are complex proteins which are very difficult to express. Blood coagulation factor IX is a vitamin K-dependent protein, and it has become a valuable biopharmaceutical in the treatment of hemophilia B. Here, we describe the techniques used to generate human cell lines producing human recombinant factor IX, as an example of complex protein, as well as in vitro characterization of this coagulation factor.

Role of crotoxin in coagulation: novel insights into anticoagulant mechanisms and impairment of inflammation-induced coagulation.

Background: Snake venom phospholipases A (svPLA) are biologically active toxins, capable of triggering and modulating a wide range of biological functions. Among the svPLAs, crotoxin (CTX) has been in the spotlight of bioprospecting research due to its role in modulating immune response and hemostasis. In the present study, novel anticoagulant mechanisms of CTX, and the modulation of inflammation-induced coagulation were investigated.

Establishment of a simple and efficient platform for car-t cell generation and expansion: from lentiviral production to in vivo studies.

Introduction: Adoptive transfer of T cells expressing a CD19-specific chimeric antigen receptor (CAR) has shown impressive response rates for the treatment of CD19 + B-cell malignancies in numerous clinical trials. The CAR molecule, which recognizes cell-surface tumor-associated antigen independently of human leukocyte antigen (HLA), is composed by one or more signaling molecules to activate genetically modified T cells for killing, proliferation, and cytokine production.

Objectives: In order to make this treatment available for a larger number of patients, we developed a simple and efficient platform to generate and expand CAR-T cells.

Production of coagulation factor VII in human cell lines Sk-Hep-1 and HKB-11.

Recombinant factor VII (rFVII) is the main therapeutic choice for hemophilia patients who have developed inhibitory antibodies against conventional treatments (FVIII and FIX). Because of the post-translational modifications, rFVII needs to be produced in mammalian cell lines. In this study, for the first time, we have shown efficient rFVII production in HepG2, Sk-Hep-1, and HKB-11 cell lines.

Approaches for recombinant human factor IX production in serum-free suspension cultures.

Objective: To establish a serum-free suspension process for production of recombinant human factor IX (rhFIX) based on the human cell line HEK 293T by evaluating two approaches: (1) serum-free suspension adaptation of previously genetic modified cells (293T-FIX); and (2) genetic modification of cells already adapted to such conditions (293T/SF-FIX).

Results: After 10 months, 293T-FIX cells had become adapted to FreeStyle 293 serum-free medium (SFM) in Erlenmeyer flasks. After 48 and 72 h of culture, 2.

Patents in therapeutic recombinant protein production using mammalian cells.

The industrial production of recombinant proteins preferentially requires the generation of stable cell lines expressing proteins in a quick, relatively facile, and a reproducible manner. Different methods are used to insert exogenous DNA into the host cell, and choosing the appropriate producing cell is of paramount importance for the efficient production and quality of the recombinant protein. This review addresses the advances in recombinant protein production in mammalian cell lines, according to key patents from the last 30 years.