Hemolysis Inhibition Profiling Service

Creative Biolabs provides world-class hemolysis inhibition profiling services to accelerate your complement therapeutic development. Gain deep, biologically relevant insights into your candidate's efficacy and mechanism of action with our robust assays. Contact us for a custom project quote.

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Why Hemolysis Inhibition Still Matters

Unlike assays that measure a single protein interaction, the hemolysis assay is a functional, systems-level readout. It doesn't just tell you if your drug binds to C5, it tells you if that binding event is sufficient to prevent the entire downstream cascade from forming the Membrane Attack Complex (MAC) and punching a hole in the cell. It captures the integrated outcome of the entire pathway, making it a crucial tool for:

Mechanism of Action (MoA) Confirmation: Verifying that your inhibitor actually works as intended in a biologically complex environment.
Potency Assessment: Providing a key metric for the concentration-dependent effectiveness of your candidate.
Comparative Analysis: Benchmarking your novel candidate against established standards-of-care.

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What We Profile

We take a source of complement, sensitize RBCs with antibodies, and trigger the classical pathway. Without an inhibitor, the cells lyse, releasing hemoglobin and turning the solution pink/red. An effective inhibitor prevents lysis, leaving the solution clear.

Pathways & Triggers

Classical Pathway (CP) - quantified as CH50 or percent hemolysis inhibition. Alternative Pathway (AP) - reported as AH50 or percent inhibition. Lectin Pathway (LP) - dependence and downstream MAC readout.

Matrices

Native human serum (normal or pooled), species-matched serum (pre-clinical models), heat-inactivated controls, and specialty lots (e.g., C3-depleted, C5-depleted) for MOA confirmation by rescue.

RBC Panels

Sheep, rabbit, chicken, and human RBC (where appropriate and ethically sourced). We tune sensitization titers, RBC density, and complement activation windows for clarity and dynamic range.

We're comfortable with inhibitor modalities: mAbs and fragments (e.g., anti-C1s, anti-C3, anti-C5), recombinant proteins (e.g., C1-INH analogs), peptides, small molecules, oligos, and formulations.

Module	Purpose	When to Use
CH50 (CP)	Global CP function & inhibitor potency	Anti-C1q/C1s/C4/C3/C5 programs; cross-check off-pathway effects
AH50 (AP)	Global AP function & inhibitor potency	Factor B/D/properdin and terminal inhibitors; safety selectivity vs. CP
LP Hemolysis	Lectin pathway selectivity	MASP-2, MBL axis; define pathway bias
Custom RBC Panel	Target relevance & sensitivity mapping	Human vs. non-human RBC response, species translations

Discuss Your Needs

Creative Biolabs Advantages

Expertise in Complement Biology

This expertise allows us to design the most appropriate profiling strategy for your specific molecule.

Comprehensive Pathway Interrogation

We offer profiling across all three complement pathways to give you a complete picture of your candidate's activity.

Standardized Protocols

We have optimized protocols for each pathway to ensure consistency and reproducibility.

Stringent Controls

Every assay includes full lysis and no lysis (no complement) controls, as well as reference standard inhibitors.

Our Typical Workflow

Creative Biolabs has armed with a powerful and standardized complement assay platform, aiming to offer the best and convenient complement related services. At present, we provide custom haemolysis inhibition assays for the rapid complement activity assessment.

The workflow of haemolysis inhibition assay. (Creative Biolabs Original) Fig. 1 Workflow of haemolysis inhibition assay.

Design Your Workflow

Sample and Material Guidance

For Your Test Articles

Provide concentration-verified stocks
Include formulation buffer for vehicle controls
Let us know if the modality is temperature-sensitive or light-sensitive

For Matrix-Dependent Questions

We'll advise exact volumes
If you want cross-donor robustness, we can source pooled serum and single-donor lots with documented complement activity

Shipping

Use cold packs (short haul) or dry ice (long haul) per modality
We provide pre-filled chain-of-custody forms

Every project is unique, and we provide extensive customization to maximize relevance.

Design Your Customization

Popular Add-On Assays

Assays. (Creative Biolabs Authorized)

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Case Studies

Case 1

Test the IC50 of Eculizumab in CH50 hemolysis assay

This assay was performed to test the IC50 of Eculizumab in CH50 hemolysis assay. Eculizumab is a potent C5 inhibitor. It inhibits the cleavage of C5 into C5a and C5b, therefor inhibits the formation of MAC. After hemolysis assay, the hemolysis was calculated and the curve was draw and finally obtained the IC50 of Eculizumab.

CH50 hemolysis assay. (Creative Biolabs Original)

Fig. 2 Hemolysis inhibitor assay.

Case 2

A hemolysis assay was carried out to investigate and compare the membrane release activity of HNH and GNH nanoparticles

A hemolysis assay was carried out to investigate and compare the membrane release activity of HNH and GNH nanoparticles at physiological pH (7.4) and endosomal pH (5.4). Both of nanoparticles were able to lyse red blood cells at pH 5.4 at different concentrations. However, the HNH showed a statistically significant higher activity than GNH at this pH. GNH nanoparticles, on the other hand, demonstrated the hemolytic activity between 6 to 30% as a function of nanoparticles concentration at pH 7.4, while the HNH nanoparticles lost their activities at this pH.

Hemolysis assay of nanoparticles. (OA Literature)

Fig. 3 Hemolysis assay of GNH and HNH Nanoparticles.^1,2

References

Alipour, Mohsen, et al. "Nano-biomimetic carriers are implicated in mechanistic evaluation of intracellular gene delivery." Scientific reports 7.1 (2017): 41507. https://doi.org/10.1038/srep41507
Distributed under Open Access license CC BY 4.0, without modification.

What Our Clients Say

"As an emerging biotechnology company, our R&D cycles and budgets are extremely tight. Your hemolysis inhibition assay service not only delivered highly accurate and reproducible data, but also provided clear and detailed reports, saving us significant time on internal validation."

— Dr. Evelyn Reed, Director of Preclinical Development

"The hemolysis inhibition analysis report they provided not only featured comprehensive data but also included profound insights and recommendations, offering invaluable reference value for our assessment of compound off-target effects and safety. Their services fully meet our standard requirements."

— Program Lead, Pharmaceutical Company

"Our research project required screening a series of natural products for hemolytic activity. Your company's service process was exceptionally streamlined, with seamless transitions from sample submission to report delivery. The final data provided robust support for our paper. We are highly satisfied with this collaboration."

— Professor Wang, Academic Research Lab

"We have been working with your company for several years, and their hemolysis inhibition assay services have been an indispensable part of our R&D process. Their customer service team is professional and friendly, always resolving our issues promptly. We highly recommend their services."

— A Long-Standing Industry Partner

Frequently Asked Questions

What species of red blood cells do you use?

We can accommodate requests for RBCs from human and other species, such as rat, mouse, rabbit, or dog, depending on your specific research needs. Please specify your requirements during your project inquiry.

What types of samples do you accept?

We accept a wide range of test articles, including but not limited to small molecules, peptides or proteins, natural product extracts, and other biologics. Samples should be provided dissolved in a suitable solvent. Please provide the exact concentration and solvent information with your submission. The required amount depends on the concentration range and the number of replicates you need. We will provide detailed sample requirements before the project begins.

What controls are standard in your studies?

Positive inhibition control (pathway-appropriate reference), negative control (vehicle/isotype), heat-inactivated serum, and—when MOA matters—component-depleted serum ± add-back. Per-plate standards normalize day-to-day drift. Outlier rules and re-run criteria are documented.

Which add-on assays pair best with hemolysis inhibition?

Frequently: C3/C4 deposition, C5b-9 quantification, C3a/C5a panels, opsonization/phagocytosis, and surface-activation models.

Can I receive preliminary updates during the experiment?

We understand your interest in the project's progress. For longer-term projects or upon your specific request, our project manager can provide you with timely updates at key milestones (e.g., upon sample receipt, after assay completion). Our goal is to maintain transparent and seamless communication.

How do I get a quote?

You can request a quote by contacting our business development team via the contact form on our website, email, or phone. Please provide as much detail as possible about your project (e.g., number of compounds, desired concentration range) so we can provide an accurate quotation.