Hypothesis:  Abzyme production is induced during pregnancy by the appearance of "foreign" factors in the blood, such as unknown DNA and/or fetal cells.

Outline of Proposed Project:                    

I. Induction of Abzyme Production:

        1. Experimentals:

                -Non-pregnant female mice

                        A. Fetal cells taken from pregnant mice will be injected intravenously into the non-                        pregnant females

                        B. DNA extracted from fetal cells will be injected into another group of experimental non-                        pregnant mice

        2. Postive Controls:

                -Pregnant mice

        3. Negative Controls:

                -Non-pregnant female mice

**Blood from each group (experimentals and positive/negative controls) will be drawn every day during the three week gestation period**

II. Purification of Abzymes:

        1. Affinity Chromatography:

                -Resins with immobilized anti-IgG and anti-IgM antibodies

            2. Gel Filtration:

                -Separation of IgG and IgM

III. Identification of Catalytic Activity:

        1. Evaluation of DNase Activity:

                -Conversion of supercoiled DNA of λ phage into circular nicking and linear forms:

                        Prepare reaction mixture of λ phage DNA + purified/separated Ab

                        Incubate at 37°C

                        Analyze by gel electrophoresis

*Example from Buneva et al., 2003*

 References:

1.  Buneva VN, Kudryavtseva AN, Gal'vita AV, Dubrovskaya VV, Khokhlova OV, Kalinina IA, Galenok VA, Nevinsky GA. 2003. Dynamics of antibody nuclease activity in blood of women during pregnancy and lactation. Biochem (Moscow) 68(8):890-900.