Hydragel IF/BJ (HR)

High resolution separation and sensitive immunological characterization of monoclonal immunoglobulins

Test presentation

Protein immunofixation is a well-established technique for the characterization of any suspected monoclonal component on serum or urine electrophoresis.

The HYDRAGEL IF/BJ (HR), when used on the HYDRASYS instrument, separates and characterizes full monoclonal immunoglobulins and Bence Jones proteins (Monoclonal Free Light Chains), in human serum or urine. The resulting patterns are longer and provide higher resolution in comparison to conventional IF and BJ kits.

The semi-automated HYDRASYS instrument offers sample application, electrophoretic migration, immunofixation and staining.

The sensitivity of the HYDRAGEL IF/BJ (HR) kit allows the detection of very small monoclonal components.

Test principle

The HYDRAGEL IF/BJ (HR) assay is based on the principle of agarose gel electrophoresis immunofixation.

The HYDRASYS system is a semi-automated multi-parameter instrument. The analytical steps include processing of the agarose gel in the following sequence:

Sample application
Electrophoretic migration
Immunofixation
Staining with acid violet dye
Destaining
Final drying

After separating serum or urine proteins according to their charge, the gel is incubated with different specific antisera targeted against:

For the IF (HR):
- Gamma- (IgG), Alpha- (IgA), Mu- (IgM) heavy chains
- Kappa (κ) and Lambda (λ) free and bound light chains

For the BJ (HR):
- Gamma- (IgG), Alpha- (IgA), Mu- (IgM) heavy chains (using the trivalent antiserum)
- Kappa (κ) and Lambda (λ) free and bound light chains
- Kappa (κ) and Lambda (λ) free light chains.

The gel is subsequently washed in order to remove unbound antisera excess prior to the final staining step. The gels are then interpreted visually. Bands present on the immunological tracks indicate the presence of monoclonal immunoglobulin and/or free light chains.

Performances

IF (HR) sensitivity:
- The detection limit for monoclonal immunoglobulin is about 60-250 mg/L for anti-IgG, anti-IgA and anti-IgM antisera. Free and bound light chain detection limit varies between 120-250 mg/L. According to the mobility of the monoclonal component and polyclonal background, the sensitivity may vary.

BJ (HR) sensitivity:
- The minimal detection limit for BJ proteins is about 12 to 25 mg/L with anti-Kappa and anti-Lambda (free and bound antisera) and as low as 50 mg/L with anti-Kappa and anti-Lambda (free light chain antisera).