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Functional estimation of mannose binding lectin associated serine protease (MBL-MASPs) in human serum
Krishana Chaitanya Gulla1, Kshitij Gupta, Krishnan Hajela
1School of Life Sciences, Devi Ahilya University, Indore, India. gullakc@yahoo.co.in
Background & Objective:
Mannose binding lectin (MBL), a C-type or Ca(2+) dependent lectin, plays a major role in lectin pathway of complement activation. MBL deficiency/insufficiency is associated with susceptibility to many infections. It is important to know the association of functional lectin levels with disease condition. Therefore, we carried out this study to develop a simple assay to estimate the functional MBL-associated serine proteases (MBL-MASPs) levels in human serum samples.
Methods:
A novel method was developed based on direct haemolysis of mannan coated human erythrocytes in autologous human serum for functional estimation of MBL and associated serine proteases (MBLMASPs complex). Functional MBL-MASPs serum levels in 75 healthy individuals was estimated. Results were compared with those obtained by ELISA based assay.
Results:
Lysis of mannan coated human RBC in autologous serum was highly specific and mediated by MBL-MASPs lectin complement pathway. Concentration of MBL-MASPs in serum of normal healthy individuals (n=75) was found to be 1.579 mug/ml (median= 1.149 mug/ml) by the haemolytic assay which was comparable to the values obtained by ELISA method.
Interpretation & Conclusion:
Our findings showed that the method developed for the estimation of functional MBL-MASPs levels in human serum is simple, cost-effective and comparable with existing ELISA method.
Insights
A new, simple, and cost-effective assay accurately measures functional mannose-binding lectin-associated serine proteases (MBL-MASPs) in human serum. This method offers a reliable alternative for assessing MBL-MASPs levels, crucial for understanding infection susceptibility.
Area of Science:
- Immunology
- Complement System Biology
Background:
- Mannose-binding lectin (MBL) is vital for the lectin complement pathway.
- MBL deficiency increases susceptibility to infections.
- Assessing functional MBL-associated serine proteases (MBL-MASPs) levels is clinically important.
Purpose of the Study:
- To develop a simple assay for estimating functional MBL-MASPs levels in human serum.
- To validate the assay's performance against established methods.
Main Methods:
- A novel direct hemolytic assay using mannan-coated erythrocytes in autologous serum.
- Functional MBL-MASPs levels were measured in 75 healthy individuals.
- Results were compared with enzyme-linked immunosorbent assay (ELISA) data.
Main Results:
- The hemolytic assay demonstrated specific MBL-MASPs-mediated lysis via the lectin complement pathway.
- Median MBL-MASPs concentration in healthy individuals was 1.149 µg/ml (mean 1.579 µg/ml).
- Hemolytic assay results closely correlated with ELISA findings.
Conclusions:
- The developed assay is simple, cost-effective, and reliable for measuring functional MBL-MASPs.
- This method provides a valuable tool for clinical and research applications.
- The assay's comparability to ELISA ensures its practical utility.
