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Printable Handouts
Navigable Slide Index
- Introduction
- Overview
- Outline
- What part of complement should you test?
- Measuring complement component levels
- Principal equivalence zone measurements
- Nephelometry
- Radial immunodiffusion (RID) principles
- Radial immunodiffusion example
- The C3 assay
- The life of C3
- C3 assay
- Complement functional assays
- Function assays
- Hemolytic functional assays
- Principle of ELISA-based functional assays
- Liposome based “CH50”
- Liposome based function assay
- Plate hemolytic CH50
- Considerations for functional assays limiting factor
- Mathematical considerations for functional assays
- Example of “Hook Effect”
- Testing for activation fragments & convertases
- Activation fragments sensitivity & specificity
- Sensitivity CH50 versus Bb
- Sensitivity CH50 versus C5a
- Split product considerations: Properties
- Split product considerations: Peak time points
- Complement genetics
- Complement genetic testing
- MFLP testing
- Immune complex testing
- Circulating immune complexes: C1q assay
- Circulating immune complexes: C3d assay
- Testing in complement specific disorders
- Complement in primary immunodeficiencies
- Complement testing for angioedema (HAE & AAE)
- HAE diagnostic test
- The ELISA based C1-INH function
- Chromogenic based C1-INH function assay
- Chromogenic assay
- Comparison of C1-INH function methods
- Method comparison
- Method comparison: Results
- Considerations with the C4 assay
- C4 on the way to the lab
- C4 assay
- C4 assay: Issue 2
- The C4 haplotypes: C4A & C4B
- Variation starting level C4
- How to handle suspect C4 results
- C4/C4d ratio assay
- Testing for paroxysmal nocturnal hemoglobinuria
- Paroxysmal nocturnal hemoglobinuria (PNH)
- Atypical hemolytic uremic syndrome (aHUS)
- aHUS testing
- More about following eculizumab
- Differences in sC5b-9 by site
- C3 glomerulopathy
- C3 glomerulopathy testing
- Pre-analytical considerations and pitfalls
- Improper storage affects split product levels: C3a
- Effect of freeze/thaw on normals vs. patients: C4a
- Does Futhan help?
- Storage effect on C1-INH function
- International efforts to standardize/improve
- Questions
Topics Covered
- The basics and foundations of common complement tests
- The current state of complement testing
- Examples of the uses of the testing for patient diagnosis
- The challenges confounding quality complement results
- The international efforts to standardize complement analysis
Links
Series:
Categories:
Therapeutic Areas:
Talk Citation
Frazer-Abel, A. (2017, June 29). Complement diagnostics [Video file]. In The Biomedical & Life Sciences Collection, Henry Stewart Talks. Retrieved October 16, 2024, from https://doi.org/10.69645/JKFK6016.Export Citation (RIS)
Publication History
Financial Disclosures
- Dr. Ashley Frazer-Abel, Consultant: Ionis Pharmaceuticals
Other Talks in the Series: The Complement System
Transcript
Please wait while the transcript is being prepared...
0:00
Welcome. I'm Ashley Frazer-Abel and I will be
speaking today on "Complement Diagnostics".
I work at the University of Colorado School of Medicine in the US where
I run a lab that specializes in complement testing.
0:14
What I'll be talking today is the testing we use in complement for patient diagnostics
to be separated from the testing that we might do for research would be more expansive.
But there's also even in complement diagnostic testing,
been quite a bit of evolving interest in this.
As you will be hearing throughout this complement series,
the number of diseases that have been recognized to be complement mediated has put
pressure on the diagnostic laboratory to be ready to diagnose those tests.
0:40
So this is the outline of what I hope to cover today.
And really to break down the complement assays into
four larger categories; complement levels,
complement functional assays, what markers
we can see for whether complement activation is occurring,
and then also complement genetics.
We'll go through some examples of complement disorders that
we would be testing in the laboratory and what tests we use for those.
But then I also want to hit on some confounding factors that we need to keep in mind
around storage and freeze thaw effects when we look at complement tests.
And then I want to close with some important international efforts to help improve and
standardize testing for complement as we go into this new area,
a new time of more interesting complement testing.
1:20
So throughout this series,
I'm sure you will see several versions of the complement pathway,
and this is the pathway version I'll be using for my lecture today.
And what I really wanna get you to get out of this,
is just that there's so many components,
and so many steps of complement that we could look at
testing diagnostically in the laboratory. So where do we start?
1:39
From my lecture, I'm gonna start with some of the simplest measurements in many ways,
and then still look at complement levels.