Biomedical subjects

Huiping Wu

Publications and source records attributed to Huiping Wu.

3 recordsLinked to original sources

PubMed2005

Process scale-up studies for protein C separation using IMAC.

Protein C (PC) is the pivotal anticoagulant and antithrombotic in the human coagulation cascade. PC deficiency can disturb the blood hemostasis and cause thrombosis, inhibiting oxygen transport to tissue, and resulting in major medical problems such as deep vein thrombosis (DVT). The current treatment can cause bleeding and other major medical problems. PC circulates in the blood as a zymogen and is only activated when and where it is needed. PC is a safe anticoagulant without harmful side effects. A combination of ion-exchange chromatography and IMAC IDA-Cu was studied for the relatively large scaled PC separation from Cohn fraction IV-1. Almost half of the active PC was recovered by using this process. In future work, we will verify the linearity of the IMAC column scale-up. This process can be used to produce PC from Cohn fraction IV-1 at large quantities and low cost to treat PC-deficient patients.

Anticoagulants↗

PubMed2003

Protein C separation from human blood plasma derivatives using low cost chromatography.

Protein C (PC) deficiency can cause thrombosis, inhibiting oxygen transport to tissue thus resulting in many complications, including death. Present treatment can cause catastrophic bleeding and other major medical problems. PC treatment has no bleeding or skin necrosis problems because it circulates in the blood as a zymogen and is only activated when and where it is needed. The vitamin K dependent (VKD) proteins are homologous proteins, making the separation of PC from plasma extremely difficult. Immobilized metal affinity chromatography (IMAC) is investigated to separate the VKD proteins to replace immunoaffinity chromatography, because of the high cost of monoclonal antibodies. An IDA-Cu column was found effective for the separation of PC from prothrombin, the most harmful contaminant. For Cohn fraction IV-1 separation, a DEAE column was found an efficient initial step, with about 25-fold PC purity increase. Following this step, an IDA-Cu column could remove many contaminants including prothrombin. The combination of DEAE and IDA-Cu resulted in PC purity increase of about 100-fold.

Chromatography, Affinity↗

PubMed2002

Chelator, metal ion and buffer studies for protein C separation.

Protein C (PC) is the pivotal anticoagulant and antithrombotic in the human coagulation cascade. PC deficiency can result in major medical problems such as deep vein thrombosis (DVT), leading to tissue oxygen deprivation. PC treatment has no bleeding or skin necrosis problems because it circulates in the blood as a zymogen and is only activated when and where it is needed. One source of PC is transgenic animal milk. The major components in the milk, such as alpha-casein, beta-casein, kappa-casein, alpha-lactalbumin and beta-lactoglobulin, are proteins that must be separated from PC. Immobilized metal affinity chromatography (IMAC) is an inexpensive separation technology with relatively high specificity, and it has great potential for difficult protein separations. After systematic studies of different chelator, metal ion and buffers, the combination of iminodiacetic acid (IDA) and Fe was found to be effective to separate PC from major milk components. alpha-Lactalbumin and beta-lactoglobulin fell through the column in the starting buffer. PC was eluted. alpha-Casein, beta-casein, kappa-casein remained bound in the column after PC elution. This technology might be applied for PC separation from transgenic animal milk. It is very important for PC production in large quantities and at low cost to treat PC-deficient patients.

Animals↗