AICDA recombinant proteins refer to engineered versions of the Activation-Induced Cytidine Deaminase (AICDA) protein, also known as AID. AICDA is a critical enzyme involved in the process of somatic hypermutation and class switch recombination of immunoglobulin genes, playing a key role in the adaptive immune response. Recombinant AICDA proteins are valuable tools for studying immune system function, developing therapeutics, and understanding the molecular mechanisms of antibody diversification.

Structure and Function

• AICDA (AID) is a cytidine deaminase enzyme that converts cytidine to uridine in DNA. This process is crucial for the diversification of antibodies through somatic hypermutation and class switch recombination.
• The enzyme is typically composed of an N-terminal domain, a central catalytic domain, and a C-terminal domain. The catalytic domain contains the active site responsible for the deamination of cytidine.
• Recombinant AICDA proteins generally include the full-length protein or specific domains necessary for its enzymatic activity and interaction with DNA or other cofactors.

Production

• Recombinant AICDA proteins are produced in expression systems such as E. coli, yeast, insect cells, or mammalian cells, depending on the need for proper folding, post-translational modifications, and activity.
• The gene encoding AICDA is cloned into an expression vector, followed by protein expression and purification using techniques like affinity chromatography. Tags such as His-tag or GST-tag are commonly used to facilitate protein purification.

Applications

• Basic Research: Recombinant AICDA proteins are used to study the mechanisms of somatic hypermutation and class switch recombination in B cells. They help in understanding how genetic diversity in antibodies is generated and regulated.
• Cancer Research: Since aberrant AICDA activity is associated with certain types of lymphomas and leukemias, recombinant AICDA proteins are used to investigate the role of AICDA in cancer development and progression.
• Therapeutic Development: AICDA is a target for therapeutic interventions in diseases where its activity is dysregulated. Recombinant AICDA proteins are used to develop and test potential inhibitors or modulators of AICDA activity.
• Diagnostics: These proteins can be used in assays to detect or measure AICDA activity or expression levels in clinical samples, aiding in the diagnosis of immunological or hematological disorders.

Validation and Quality Control

• Validation of recombinant AICDA proteins involves confirming their identity, purity, and functionality using methods such as SDS-PAGE, Western blotting, and mass spectrometry.
• Functional assays, such as enzymatic activity assays or DNA mutagenesis assays, are performed to ensure that the recombinant proteins are biologically active and capable of performing their intended functions.