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Multiplex Assay Kit for Mannose Binding Lectin (MBL) ,etc. by FLIA (Flow Luminescence Immunoassay)

MBL2; COLEC1; HSMBPC; MBP1; MBP; Collectin-1; Mannose-binding protein C; Mannan Binding Protein; Mannose-Binding Lectin(Protein C)2,Soluble(Opsonic Defect)

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Mannose Binding Lectin (MBL) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Mannose Binding Lectin (MBL) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Mannose Binding Lectin (MBL) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Mannose Binding Lectin (MBL) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 88-96 92
EDTA plasma(n=5) 81-99 90
heparin plasma(n=5) 82-99 95
sodium citrate plasma(n=5) 78-99 87

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Mannose Binding Lectin (MBL) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Mannose Binding Lectin (MBL) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Mannose Binding Lectin (MBL) ,etc. by FLIA (Flow Luminescence Immunoassay) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 78-89% 94-101% 88-102% 96-104%
EDTA plasma(n=5) 79-104% 96-104% 84-94% 81-103%
heparin plasma(n=5) 80-101% 89-96% 80-91% 82-90%
sodium citrate plasma(n=5) 78-89% 83-103% 89-97% 92-99%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:MBL) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
American Journal of Nephrology Analysis of the Urine Proteome of Human Contrast-Induced Kidney Injury Using Two-Dimensional Fluorescence Differential Gel Electrophoresis/Matrix-Assisted Laser Desorption Time-of-Flight Mass Spectrometry/Liquid Chromatography Mass Spectrometry. Karger: 000255439
Current Eye Research Serum Levels and H/L Gene Polymorphism of Mannose-Binding Lectin in Primary Open Angle Glaucoma Ingenta: art00007
Journal of Clinical Immunology Complement activation contributes to the injury and outcome of kidney in human anti-glomerular basement membrane disease. PubMed: 22941511
Clinical and Experimental Immunology Urinary mannose-binding lectin is a biomarker for predicting the progression of immunoglobulin (Ig)A nephropathy PubMed: PMC3406374
Clin J Am Soc Nephrol. Alternative Complement Pathway Activation Products in Urine and Kidneys of Patients with ANCA-Associated GN Pubmed: 24115193
ACTA TROPICA Novel findings on the role of ficolins and colectins in the innate response against Leishmania braziliensis Pubmed: 32827456
PLoS One Complement activation profile of patients with primary focal segmental glomerulosclerosis Pubmed: 32569286
LIFE SCIENCES The lectin pathway of complement and the initial recognition of Leishmania infantum promastigotes 34242658
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