Anti-HLA Class II Control

Monoclonal IgG1 positive control for class II Luminex® (LABScreen® or LIFECODES HLA)

IgG1 is the most abundant IgG in serum (66%). IgG1 antibodies strongly activate complement and exhibit strong cellular effector function.

Anti-HLA Class II Control is a chimeric human IgG1 monoclonal antibody specifically designed for studying detection of human IgG antibodies to HLA class II. Its constant regions are from human IgG1 (i.e. human IgG1 heavy chains and kappa light chains) and its variable regions are derived from the murine monoclonal anti-HLA class II antibody F3.3 [1].

Anti-HLA Class II Control can be used as an IgG positive control for IgG anti-HLA antibody detection assays, including LABScreen®, LIFECODES HLA and C1qScreen™.

Anti-HLA Class II Control has been validated in HLA Class II LABScreen® Mixed and LABScreen® Single-Antigen. It reacts with all HLA-DP, DR and DQ2 antigens but not with DQ1, DQ3 & DQ4 [2].

The anti-HLA Class II Control also binds strongly to recombinant C1q, using the C1qScreen™ (see figures).

References:

1. Elsässer D. et al., 1996. HLA class II as potential target antigen on malignant B cells for therapy with bispecific antibodies in combination with granulocyte colony-stimulating factor. Blood. 87(9):3803-12.
2. Congy-Jolivet N et al. 2013. Production and characterization of chimeric anti-HLA monoclonal antibodies targeting public epitopes as tools for standardizations of the anti-HLA antibody detection. J Immunol Methods. 390(1-2):41-51

Figures

Typical fluorescent dose-response obtained from serial dilution of Anti-HLA Class II Control (hIgG1) antibody (μg/ml) in a LABScreen® Mixed assay using PE-conjugated anti-human IgG as label. The curve has been plotted with average MFI values for the class II beads (minimal standard deviation as each bead gave a very similar value). The signal for Class I beads (not shown) was at background level.

Fluorescent (MFI) dose-response curves obtained for binding of anti-HLA class II control to LABScreen® Single Antigen beads. Each curve represents a specific HLA antigenic bead. The antigens that gave the lowest values are indicated on the right. As observed in the graph, interaction of the anti-HLA class II control with HLA-DQB1*03, 04, 05 and 06 is weak or absent. background level.

Average fluorescent intensity obtained at 1 μg/ml of Anti-HLA Class II Control (hIgG1) on LABScreen® Mixed and C1qScreen™. The data are average MFIs of the class II beads. Human IgG2 (hIgG2) and hIgG4 antibodies were produced separately to assess sensitivity. The class I beads signal (not shown) was at background level.

Specifications

Working concentration: 0.2 ng/ml – 2 µg/ml

Specificity: HLA-DP, DR, and DQ2. It does not react with DQ1, DQ3 or DQ4 antigens.

Isotype: Human IgG1, kappa

Formulation: 0.2 µm filtered solution in PBS with 5% saccharose.

Applications: Positive controls for all anti-HLA antibody detection assays, including:

• multiplex bead array assays using Luminex® technology (LABScreen®, LIFECODES HLA, C1qScreen™);
• ELISA;
• indirect immunofluorescence techniques (flow cytometry);
• complement-dependent lymphocytotoxicity.

Contents

• 100 µg sterile Anti-HLA Class II hIgG1 antibody, provided azide-free and lyophilized.

Shipped at room temperature.

Store the antibody at -20°C.

Lyophilized product is stable 24 months at -20°C