1887

Chapter 14 : An Enzyme-Linked Immunosorbent Assay-Based Method for Functional Analysis of the Three Pathways of the Complement System

MyBook is a cheap paperback edition of the original book and will be sold at uniform, low price.

Preview this chapter:
Zoom in
Zoomout

An Enzyme-Linked Immunosorbent Assay-Based Method for Functional Analysis of the Three Pathways of the Complement System, Page 1 of 2

| /docserver/preview/fulltext/10.1128/9781555815905/9781555813642_Chap14-1.gif /docserver/preview/fulltext/10.1128/9781555815905/9781555813642_Chap14-2.gif

Abstract:

The complement system has a crucial role in innate immune defense against invading microorganisms and can be activated by the classical pathway (CP), the alternative pathway (AP), and the mannose-binding lectin (MBL) pathway (MP). The CP is activated by binding of C1q to, e.g., immunoglobulins present on microorganisms or by direct binding to apoptotic cells. The AP can be directly activated by invading microorganisms. The MP is also directly activated, via carbohydrate moieties present on the surface of invading microbes. For assessment of the functional activity of the classical and alternative pathways, hemolysis of erythrocytes by complement activation via either the CP (CH50) or the AP (AP50) is used in most laboratories. The methods to assess pathway activity of the CP, AP, and MP are enzyme-linked immunosorbent assay (ELISA)-based. All three assays are delivered as one ELISA system. Blood samples are to be collected under sterile conditions in red-top tubes, without serum separator. A positive control is provided in the kit. Frozen sera is partially thawed by briefly placing them in a 37№ C water bath with gentle mixing. With the combined assay, the functional activity of the three pathways of complement activation can be assessed. With the use of combined ELISA system, the three pathways for complement activation can be assessed at the same time.

Citation: Seelen M, Roos A, Wieslander J, Daha M. 2006. An Enzyme-Linked Immunosorbent Assay-Based Method for Functional Analysis of the Three Pathways of the Complement System, p 124-127. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch14

Key Concept Ranking

Complement System
0.64845943
Complement Components
0.5478034
Enzyme-Linked Immunosorbent Assay
0.54528147
0.64845943
Highlighted Text: Show | Hide
Loading full text...

Full text loading...

References

/content/book/10.1128/9781555815905.ch14
1. Aderem, A., and , D. M. Underhill. 1999. Mechanisms of phagocytosis in macrophages. Annu. Rev. Immunol. 17:593623.
2. Fredrikson, G. N.,, L. Truedsson, and , A. G. Sjoholm. 1993. New procedure for the detection of complement deficiency by ELISA. Analysis of activation pathways and circumvention of rheumatoid factor influence. J. Immunol. Methods 166:263270.
3. Ikeda, K.,, T. Sannoh,, N. Kawasaki,, T. Kawasaki, and , I. Yamashina. 1987. Serum lectin with known structure activates complement through the classical pathway. J. Biol. Chem. 262:74517454.
4. Kuipers, S.,, P. C. Aerts,, A. G. Sjoholm,, T. Harmsen, and , H. van Dijk. 2002. A hemolytic assay for the estimation of functional mannose-binding lectin levels in human serum. J. Immunol. Methods 268:149157.
5. Minchinton, R. M.,, M. M. Dean,, T. R. Clark,, S. Heatley, and , C. G. Mullighan. 2002. Analysis of the relationship between mannose-binding lectin (MBL) genotype, MBL levels and function in an Australian blood donor population. Scand. J. Immunol. 56:630641.
6. Petersen, S. V.,, S. Thiel,, L. Jensen,, R. Steffensen, and , J. C. Jensenius. 2001. An assay for the mannanbinding lectin pathway of complement activation. J. Immunol. Methods 257:107116.
7. Roos, A.,, L. H. Bouwman,, J. Munoz,, T. Zuiverloon,, M. C. Faber-Krol,, F. C. Fallaux-van den Houten,, N. Klar-Mohamad,, C. E. Hack,, M. G. Tilanus, and , M. R. Daha. 2003. Functional characterization of the lectin pathway of complement in human serum. Mol. Immunol. 39:655668.
8. Walport, M. J. 2001. Complement. First of two parts. N. Engl. J. Med. 344:10581066.

Tables

Generic image for table
TABLE 1

Diagram for placement of the diluted sera from patients and controls in the assay plate

Citation: Seelen M, Roos A, Wieslander J, Daha M. 2006. An Enzyme-Linked Immunosorbent Assay-Based Method for Functional Analysis of the Three Pathways of the Complement System, p 124-127. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch14
Generic image for table
TABLE 2

Determination of possible complement deficiency by combination of the test results from the three assays

Citation: Seelen M, Roos A, Wieslander J, Daha M. 2006. An Enzyme-Linked Immunosorbent Assay-Based Method for Functional Analysis of the Three Pathways of the Complement System, p 124-127. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch14

This is a required field
Please enter a valid email address
Please check the format of the address you have entered.
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error