If you want your Chemiluminescence CLIA kit to give accurate and reliable results, there are a few important things to pay attention to. Here’s a breakdown of the key factors:
What To Consider When Improving Performance?
1. Make Sure Magnetic Beads Are Mixed Properly
CLIA tests often use magnetic beads to capture the target substance. If these beads aren’t mixed well inside the machine before testing, your first result might be way too high or too low—this is called the “first sample effect.”
Why does this happen? If the beads are unevenly spread, the test won’t have the right amount of particles, which affects accuracy.
Things that can affect this include:
- The size of the beads
- How they’re prepared
- The buffer (liquid) they’re stored in
| Tip: It’s best to catch and fix this issue early in the development phase. |
2. Choose Between a One-Step or Two-Step Test
You can design your CLIA test in one or two steps:
- One-step tests are faster.
- Two-step tests include a wash in the middle, which helps reduce non-specific binding (unwanted interactions).
If your test is picking up too much background noise or false positives, a two-step version might be better.
3. Reduce Non-Specific Binding
Sometimes the test reacts with things it’s not supposed to—like certain antibodies in human blood (e.g., HAMA or heterophilic antibodies).
To prevent this:
- You can add extra antibodies (like polyclonal and mouse monoclonal antibodies) to the assay buffer.
- These help block unwanted reactions and improve test accuracy.
4. Test Reagent Stability
Your test reagents need to stay stable during storage and shipping. That means they should still work well even after being on the shelf for a while.
To test this:
- Reagents are often kept at high temperatures (like 37–46°C) for a few days or weeks.
- If performance drops during this time, it may mean the reagents won’t last long in real-world conditions.
Real-time tests and shipping simulations are still needed, but these accelerated tests help spot issues early.
5. Use Lyophilization (Freeze-Drying) if Needed
If keeping reagents stable is a problem, some companies freeze-dry their reagents. This helps them last longer without refrigeration.
Things To Consider When Scaling Up a Chemiluminescent Immunoassay (CLIA)
1. Set Control Checkpoints
As you scale up, it’s helpful to set control points—checkpoints in your process where you test for inconsistencies or problems.
These checkpoints help you quickly spot issues with materials or steps in the process before they become bigger problems.
2. Use the Same Equipment
Try to use the same type of instruments or machines for both small and large batches. It helps keep results consistent. Further, it reduces surprises when you move to larger production.
3. Scale Key Reagents
Important ingredients in your test—like the CLIA-labeled proteins—need to be available. You need them in larger amounts when scaling up.
Double-check that you can reliably produce or source enough of these critical reagents. It should be done before you expand your full process.
4. Prepare for Bigger Needs
Scaling up means everything gets bigger—not just the batch size. You’ll also need to increase:
- Storage space
- Temperature-controlled areas
- Magnetic separators
- Lab equipment and materials
Make sure your facility and supplies can handle the larger scale without affecting quality.
Planning helps make the transition to large-scale production smooth and reliable.