Signal enhancer BAP-1

The signal booster BAP-1 has a unique molecular structure that gives it multifunctional properties in immunodiagnostics. The core functions are: preventing particle aggregation, enhancing reagent compatibility, improving luminescence efficiency, protecting enzyme activity, reducing non-specific adsorption and inhibiting spontaneous oxidation.

Specific dosage, mechanism and expected effect of signal enhancer BAP-1 in immunodiagnosis

Chemiluminescence

· Recommended concentration:

o Conventional stability: 0.1-0.5% (w/v);

o hydrophobic matter solubilization: 0.5-1% (w/v).

· Core role:

o Disperse luminescent reagents (such as luminol, acridinium ester) to prevent aggregation;

o Protect enzyme (such as HRP, ALP) activity, reduce non-specific adsorption;

o micelles wrap hydrophobic molecules, enhance the local reaction concentration, improve the luminous efficiency.

· Expected effect:

The intensity of the luminous signal is increased by 2-5 times, and the background noise is reduced by 30-50%;

o enzyme activity retention rate >90%, reagent stability extended to more than 24 hours.

Colloidal Gold (Colloidal Gold)

· Recommended concentration:

o Pretreatment stability: 0.1-0.5% (w/v);

o Coupling buffer: 0.05-0.1% (w/v).

· Core role:

o Prevent gold particles from aggregating through steric hindrance;

o Reduce non-specific binding during antibody coupling;

o Optimize the surface charge and enhance the coupling efficiency.

· Expected effect:

o uniform dispersion of colloidal gold (PDI <0.2);

The antibody coupling efficiency is improved by 20-30%, and the detection sensitivity is pg/mL.

Quantum Dots (QDs)

· Recommended concentration:

o Pretreatment: 0.5-2% (w/v);

o Long-term storage: 0.5-1% (w/v).

· Core role:

o Prevent aggregation by providing steric hindrance;

o Protect quantum dot fluorescence from environmental quenching.

· Expected effect:

o Stability of quantum dot aqueous solution >1 month (4℃ away from light);

o Fluorescence quantum yield retention rate >80%, particle size distribution unimodal (DLS verification).

Time-Resolved Fluorescence (TRF)

· Recommended concentration:

o Rare earth fluorescence microsphere stabilization: 0.05-0.2% (w/v);

o Reduced background: 0.05-0.1% (w/v).

· Core role:

o Reduce the non-specific adsorption of microspheres or markers;

o Prolong fluorescence lifetime stability, inhibit environmental interference;

o Enhance the signal-to-noise ratio of the probe in complex samples, such as serum.

· Expected effect:

The background signal is reduced by 50-70%, and the signal-to-noise ratio (SNR) is increased by 3-5 times;

o Time-resolved signal stability extended to 72 hours.

Microfluidic immunoassay chip

· System: PDMS chip + antibody modified channel + serum sample detection

· Formula:

o Chip pretreatment: 1% signal enhancer BAP-1 was injected for 30 minutes, and PBS was washed;

o Mobile phase: 0.2% coupling enhancer BAP-1 + 0.1% BSA (pH 7.4).

· Effect:

o Non-specific adsorption reduced by 70%, detection limit up to 0.1 pg/mL;

o Chip reuse increased from 3 to 15 times.

Droplet microfluidic single cell analysis

· System: water phase (including cells) + oil phase (fluorocarbon oil + surfactant)

· Formula:

o Aqueous phase: 0.1% signal enhancer BAP-1 + cell suspension;

o Oil phase: 2% PFPE-PEG surfactant.

· Effect:

Droplet generation rate 10,000/ min, single cell encapsulation rate > 85%;

o Cell activity maintained >90% (24 hours culture).

Temperature-responsive drug controlled release chip

· System: Signal enhancer BAP-1 gel coated with doxorubicin (DOX) + heating module

· Formula:

o Gel phase: 2% signal enhancer BAP-1 + 1 mg/mL DOX (liquid loading at 4℃);

Controlled release condition: local heating to 37℃ gelation, continuous release.

· Effect:

o Drug release time extended to 48 hours;

o Tumor cell killing rate increased by 40% (compared to direct injection).

Rapid qPCR

Concentration range

· Conventional rapid qPCR: 0.05%-0.2% (w/v), it is recommended to optimize from 0.1%.

· Ultra-fast qPCR (<30 minutes) : 0.2%-0.5% can be tried, but the risk of inhibition of enzyme activity needs to be verified.

Optimization step

· Gradient test: 0%, 0.1%, 0.2%, 0.5% concentrations were set, and Ct values and amplification efficiency of low-concentration templates (e.g. 10 copies/μL) were compared.

Compatibility verification: Ensure compatibility with fluorescent dyes (such as SYBR Green), reverse transcriptase (one-step RT-qPCR), and avoid quenching or background elevation.

· Core role:

o Reduce non-specific adsorption

Rapid qPCR reaction time is short, the temperature changes quickly, and the contact time between reagent and tube wall is reduced, but the high frequency of rising and cooling may lead to local concentration or adsorption of reagent.

The signal enhancer BAP-1 reduces the non-specific adsorption of DNA/ enzyme through the coated reaction tube wall, especially for the capture efficiency of low concentration templates (such as trace pathogen nucleic acid or single-cell RNA).

o Stabilization of heat-sensitive components

In rapid qPCR, components such as Taq enzymes may be partially inactivated by violent temperature fluctuations.

Signal enhancer BAP-1 stabilizes enzyme structure through hydrophobic interaction, maintains activity at high temperature, and reduces the loss of amplification efficiency.

o Suppress bubble interference

Rapid rising and cooling is easy to produce bubbles, which interferes with the acquisition of light signals (such as affecting the reading of fluorescence values).

Signal enhancer BAP-1 reduces liquid surface tension, reduces bubble formation and ensures fluorescence detection stability.

Concentration optimization strategy

· Gradient experiment: start from a low concentration (such as 0.05%) and gradually increase to the target effect;

· Compatibility test: Verify the compatibility of the signal booster BAP-1 with system components (such as enzymes, antibodies, buffers).

Combined strategy:

Firstly, the nanoparticles were stabilized by signal enhancer BAP-1.

After the antibody was conjugated, it was blocked with BSA.

A low concentration of the signal enhancer BAP-1 (0.1%) is retained in the end product to maintain long-term stability.

Recommended sealer combination:

1% signal enhancer BAP-1 + 0.1% Tween-20; Closure for in vitro testing with low background requirements

2% signal enhancer BAP-1 + 0.5% casein; Suitable for sealing without animal origin ingredients

0.1% signal enhancer BAP-1+1% BSA; Suitable for high sensitivity detection closure

When using signal enhancer BAP-1 in vitro diagnostic (IVD) reagents, the following key considerations and solutions need to be considered:

Concentration optimization and stability

Concentration control

· Risk:

o concentration too high (>1%) : may cause micelles to wrap active molecules (such as antibodies, enzymes), inhibit reaction efficiency;

o concentration too low (<0.05%) : unable to effectively disperse particles or inhibit non-specific adsorption.

· Suggestions:

o Through the gradient experiment (0.05%, 0.1%, 0.2%, 0.5%, 1%) optimization, select the lowest effective concentration;

o Conventional starting concentrations: 0.1-0.2% (chemiluminescent/colloidal gold systems) or 0.5-1% (quantum dots/hydrophobic systems).

Biocompatibility and interference

1. Influence on biological activity

· Risk:

The signal enhancer BAP-1 may partially cover the active site of the antibody/enzyme, reducing the coupling efficiency or catalytic activity.

· Suggestions:

o Verify the activity of key biomolecules, such as HRP enzyme activity in ELISA (TMB color rendering);

o Preferentially add the signal booster BAP-1 after coupling (e.g. after colloidal gold labeling antibody).

2. Sample compatibility

· Risk:

High concentrations of the signal enhancer BAP-1 may interfere with protein interactions in serum/plasma samples, resulting in false negatives/positives.

· Suggestions:

o Clinical validation: Testing of reagent properties in real samples (such as patient serum);

o Add competitive sealer: 1% BSA or 0.5% casein in combination.

Physicochemical compatibility

1. Temperature sensitivity

· Risk:

The signal booster BAP-1 may form a gel at >20℃, blocking microfluidic channels or affecting liquid flow.

· Suggestions:

o Control reaction temperature: keep 15-25 ℃ (avoid gelation);

Pre-experimental testing: observation of reagent fluidity at different temperatures (e.g. dynamic light scattering monitoring of particle size).

2. Interaction with other reagents

· Risk:

o combined with ionic surfactants (such as SDS, CTAB) to destroy the micellar structure;

o In combination with high salt buffers (such as PBS) to reduce steric hindrance.

· Suggestions:

o Avoid active combination with ionic surface, preferentially select Tween-20 (non-ionic) auxiliary dispersion;

Salt concentration control: ≤ 0.15M NaCl (colloidal gold system) or ≤ 0.1M (quantum dot system).