Monoclonal antibodies to human coagulation factor V and factor Va

Area of Science:

• Immunology
• Hematology
• Biochemistry

Background:

• Human factor V is a crucial protein in the blood coagulation cascade.
• Understanding factor V activation and its components is vital for hemostasis research.
• Monoclonal antibodies are powerful tools for dissecting protein structure and function.

Purpose of the Study:

• To generate and characterize monoclonal antibodies against human factor V and its activated form, factor Va.
• To utilize these antibodies to investigate the subunit composition and activation of factor Va.
• To identify the epitope recognized by a specific anti-factor V antibody (alpha HFV-1).

Main Methods:

• Immunization of BALB/c mice with human factor V (procofactor and factor Va).
• Hybridoma technology for antibody production, followed by radioimmunoassay screening.
• Subcloning of positive hybridomas and ascites production for antibody characterization.
• Affinity chromatography using an anti-factor V antibody (alpha HFV-1) for factor Va subunit analysis.

Main Results:

• Successfully generated hybridoma cell lines producing antibodies specific to factor V and/or factor Va.
• Characterized antibodies showing differential reactivity with factor V and factor Va.
• Demonstrated that human factor Va is composed of two subunits (93,000 and 70,000 molecular weight).
• Identified the epitope for the alpha HFV-1 antibody resides on the 70,000 molecular weight subunit of factor Va.

Conclusions:

• Monoclonal antibodies provide valuable tools for studying coagulation factors.
• Human factor Va is a heterodimeric protein.
• The alpha HFV-1 antibody recognizes an epitope on the 70 kDa subunit of factor Va, aiding in structural studies.