Product information

Diagnostic reagents

Additives for diagnostic reagents

NOF is developing Biolipidure® series and LIPIDURE®-SF series as additives for in vitro diagnostics.

Function of LIPIDURE®

As additives for diagnostics, Biolipidure® has multiple functions such as suppression of non-specific adsorption of protein, stabilization of protein, and sensitization, all of which are required for developing diagnostic reagents.

Basic function of Biolipidure®

The table below shows basic functions of Biolipidure® as additives for diagnostic reagents.
The Biolipidure® series has great variety of polymers having different physical properties, which is derived from rich variation of secondary monomers copolymerized with MPC.

Biolipidure®－ Side chain property Blocking effect Stabilizing effect Sensitizing effect Improvement of reproducibility and accuracy Solubilizing effect
100 series Hydrophilic 　 ○ ◎ ◎ 　
200 series Hydrophobic ◎ ◎ ◎ ◎ ◎
400 series Anion 　 　 ◎ ○ 　
500 series Cation 　 　 ○ 　 　
700 series Hydrogen bond 　 　 ○ 　 　
800 series Hydrophilic
Hydrophobic		 ◎ ◎ ○ ◎ 　
1000 series Hydrophobic ◎ ◎ ◎ ◎ ◎
1200 series Hydrophobic ○ ○ ○ 　 ○
1300 series Hydrophobic ○ ○ ○ 　 ○

◎ : Best
○ : Good

Index

Products
-[Polymer type] Biolipidure® series
-[Zwitterionic surfactant type] LIPIDURE®-SF08
Basic properties of LIPIDURE®
-Blocking effect
-Protein-stabilization effect
-Sensitizing effect
Application of Biolipidure®
-Latex agglutination
-Immunochromatography
-ELISA / CLEIA

Products

[Polymer type] Biolipidure® series

Product nameCharacteristicsForm
Biolipidure®-103Hydrophilic side chain5 wt% aqueous solution
Biolipidure®-203Hydrophobic side chain5 wt% aqueous solution
Biolipidure®-206Hydrophobic side chain5 wt% aqueous solution
Biolipidure®-405Anionic side chain5 wt% aqueous solution
Biolipidure®-502Cationic side chain5 wt% aqueous solution
Biolipidure®-702Hydrogen bonding side chain5 wt% aqueous solution
Biolipidure®-802Hydrophobic/hydrogen bonding side chain5 wt% aqueous solution
Biolipidure®-1002Hydrophobic side chain5 wt% aqueous solution
Biolipidure®-1201Hydrophobic side chain5 wt% aqueous solution
Biolipidure®-1301Hydrophobic side chain5 wt% aqueous solution

[Zwitterionic surfactant type] LIPIDURE®-SF08

The LIPIDURE®-SF08 is a low molecule additive for diagnostic reagents.

Product nameCMC [wt%]Form
LIPIDURE®-SF080.15 wt% aqueous solution

Basic performance of Biolipidure®

Blocking effect

Biolipidure® suppresses non-specific adsorption of biomolecule. NOF provides a set of products that can coat the surface of plastic, metal or other various materials.
Biolipidure® consists of wholly synthesized polymers, so it is free from biohazardous risk and has little lot-to-lot variation, making Biolipidure® the best blocking agent for diagnostic reagents.

Example 1: Blocking microplates

After a microplate was treated with Biolipidure®, the amount of non-specific adsorption of enzyme-labeled antibody was measured. The amount of non-specific adsorption of enzyme-labeled antibody was significantly reduced, proving that Biolipidure® provides excellent blocking effect.

[Test procedure]

  1. 200 µL of LIPIDURE® (diluted 10-fold by PBS) was applied to a 96-well plate.
  2. The plate was aspirated and dried overnight.
  3. 100 µL of 24,000-fold diluted HRP-IgG was added.
  4. After 1 hr incubation at a room temperature, the plate was washed with PBS-T.
  5. TMB colorimetric substrate (KPL) was applied and the non-specific adsorption of HRP-IgG was measured.

Example 2: Blocking magnetic beads

After magnetic beads was treated with Biolipidure®, the amount of non-specific adsorption of enzyme-labeled antibody was measured.
The amount of non-specific adsorption of enzyme-labeled antibody was significantly reduced, showing that Biolipidure® provides excellent blocking effect.

RLU: Relative Light Unit

[Test procedure]

  1. 170µg/mL of magnetic beads suspension was added to the same amount of Biolipidure® diluted 5-fold by TBS.
  2. After 1 hr of incubation at a room temperature, the beads was washed with TBS.
  3. 10 ng of AP-IgG was added.
  4. After 1 hr of incubation at a room temperature, the beads was washed with TBS–T.
  5. Chemiluminescent substrate was applied and non-specific adsorption of AP-IgG was measured.

Example 3: Blocking magnetic beads (Antibody solution)

The same protein-blocking effect was also achieved when Biolipidure® was added to an antibody solution. Biolipidure® was added to a diluted solution of enzyme-labeled antibody, and the amount of non-specific adsorption to magnetic beads was measured.
The amount of non-specific adsorption of enzyme-labeled antibody was significantly reduced, proving that LIPIDURE® provides excellent blocking effect.
* Biolipidure® does not affect antibody titer.

RLU: Relative Light Unit

[Test procedure]

  1. 0.1 times volume of Biolipidure® added to 100 ng/mL of AP-IgG solution.
  2. 100 µL of the AP-IgG solution containing Biolipidure® was added to 8.6 µg of magnetic beads.
  3. After 1 hr of incubation at a room temperature, the beads was washed with TBS-T.
  4. Chemiluminescent substrate was applied and non-specific adsorption of AP-IgG was measured.

Protein-stabilization effect

Biolipidure® stabilizes proteins, probably because the interaction between Biolipidure® and proteins maintains the native state structure of the proteins.

Example 1: Stabilization of antibodies (Solid phase)

The stability of immobilized capture antibody was improved by treating the surface od the microplate by Biolipidure®. This treatment also reduces the non-specific adsorption of proteins applied subsequently.
After the capture-antibody-immobilized microplate was coated with Biolipidure® and stored at 25°C, the residual antibody titer was measured using ELISA (sandwich method).
By using Biolipidure®, the stability of antibody was improved, that is, alsmost 100% of antibody titer was maintained for more than 15 days.

Relative activity: Measured using activity of Day 0 as 100%.

[Test procedure]

  1. Anti-mouse IgG antibody (capture antibody) was immobilized on a microplate.
  2. After 1 hr of incubation at a room temperature, the plate was washed with PBS.
  3. 200 µL of LIPIDURE® diluted 10-fold by PBS was added.
  4. After 1 hr of incubation at a room temperature, the plate was washed with PBS.
  5. The microplate was stored at 25°C in desiccator.
  6. Mouse IgG (antigen) was added.
  7. After 1 hr of incubation at a room temperature, the plate was washed with PBS-T.
  8. HRP labeled anti-mouse IgG antibody was added.
  9. After 1 hr of incubation at a room temperature, the plate was washed with PBS-T.
  10. Colorimetric substrate (TMB) was added and the antibody titer of the capture antibody was measured.

Example 2: Stabilization of antibodies (Liquid phase)

LIPIDURE®-SF provides stabilization effect when added to a diluted solution of enzyme-labeled antibody.
LIPIDURE®-SF was added to a diluted solution of enzyme-labeled antibody and residual enzyme activity was measured after storage at 4°C.
By adding LIPIDURE®-SF, the stability of enzyme-labeled antibody was markedly improved and 100% of enzyme activity was maintained for 60 days.
* LIPIDURE®-SF does not affect antibody titer.

Relative activity: Measured using activity of Day 0 as 100%.

[Test procedure]

  1. 0.02 times volume of LIPIDURE®-SF08 was added to 10,000-fold diluted solution of enzyme-labeled antibody.
  2. The solution was stored at 4°C.
  3. The residual activity of labelled enzyme was measured with colorimetric substrate (TMB).

Sensitizing effect

BIOLIPIDURE® exhibits sensitizing effect in immunoassay systems such as latex agglutination or immunochromatography, most likely caused by the improved dispersion of latex particle or gold colloid.

Example 1: Sensitizing effect of Biolipidure® in latex agglutination test

Biolipidure® provides excellent sensitizing effect in latex agglutination test.

[Test procedure]

CRP: C-reactive protein)
(Detection was performed using an automated analyzer.)

Example 2: Sensitizing effect of Biolipidure® in lateral flow test (immunochromatography)

Biolipidure® provides excellent sensitizing effect in lateral flow test (immunochromatography).

[Test procedure]

(hCG: human chorionic gonadotropin)
(The immunochromatography kit is manufactured in our laboratory.)

Application examples using Biolipidure®

Latex agglutination test

Biolipidure® provides excellent sensitizing effect in latex agglutination test.

[Test procedure]

(CRP: C-reactive protein)
(Detection was performed using an automated analyzer.)

Lateral flow test (Immunochromatography)

Biolipidure®, especially Biolipidure-40X, provides improved sensitizing effect in lateral flow test (immunochromatography).

Dengue virus kit

[Test procedure]

Adenovirus kit

[Test procedure]

ELISA / CLEIA

Biolipidure® provides sensitizing effect in sandwich type Chemiluminescent Enzyme Immunoassay (CLEIA).

[Test procedure]

  1. Mouse anti-Human IgG antibody was immobilized on a microplate.
  2. After 1 hr of incubation at a room temperature, the plate was washed with TBS.
  3. Biolipidure® solution (0.1 times volume) was added.
  4. The plate was incubated for 1 hr at a room temperature.
  5. Human IgG (antigen) standard was added.
  6. After 1 hr of incubation at a room temperature, the plate was washed with TBS-T.
  7. AP labeled anti-Human IgG antibody was added.
  8. After 1 hr of incubation at a room temperature, the plate was washed with TBS-T.
  9. Chemiluminescent substrate was added and calibration curve for Human IgG concentration was prepared.

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