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Your Position: Start > Anti-Idiotypic Antibody Development Service

Anti-Idiotypic Antibody Development Service

Background

Anti-idiotypic antibodies are antibodies that target the variable region of another antibody to produce specific binding. These antibodies are widely used in drug development: they can be used as an important reference standard for immunogenicity analysis or can be developed to specifically detect antibody drug levels for pharmacokinetic studies.

ACROBiosystems is committed to providing high quality key reagents and related services required in the development of targeted therapeutic drugs. We provide a series of anti-idiotypic antibodies with high affinity, high specificity, and high sensitivity. To meet the more unique needs of our customers, we also provide a one-stop service, starting from antigen preparation to monoclonal / polyclonal anti-idiotype antibody and pharmacokinetic / immunogenicity test kit development.

Advantages


Hardware Strength

Through the international AAALAC certification, we can standardize the management and use of animals to ensure the quality of antibodies.


Efficient Project Management

Dedicated one-on-one service with our project team with fast response and follow-up.


High Quality Materials

Key reagents for PK/ADA analysis are provided for a variety of different types of therapeutic drugs including monoclonal antibodies, bispecific antibodies, ADCs, and CAR-T).


Multifunctional Service

Services starting from antigen preparation to monoclonal / polyclonal anti-idiotype antibody development and PK/immunogenicity testing kit development.


Compliant with Regulatory Guidelines

We ensure that our PK/ADA test kits are delivered with sensitivity that meets regulatory requirements.

Services
Monoclonal anti-idiotypic antibodies preparation service mouse

We provide custom development of anti-idiotypic antibodies that are either antigen-neutralizing, non-neutralizing or drug target compound types. Depending on your application, these monoclonal anti-idiotypic antibodies can be used in the detection of free, bound, or total drug concentration in pharmacokinetic studies.

ada_2
Antigen blocking type(Neutralizing)
Paratope specific
Block antigen-antibody binding
Detection of free antibody drugs
Detection of neutralizing antibodies
ada_3
Antigen non-blocking type (Non-neutralizing)
Paratope non-specific
Not block antigen-antibody binding
Detection of total antibody drugs (free, semi-bound, bound)
ada_4
Drug target compound type
Specific binding drug target complex
Not block antigen- antibody binding
Specific detection of bound antibody drugs
Polyclonal anti-idiotypic antibody preparation service mouse

For immunogenicity studies, we provide high-affinity and specific polyclonal anti-idiotypic antibody development services. These highly sensitive and diverse polyclonal anti-idiotypic antibodies can be used as a positive reference in immunogenicity studies, simulating the production of ADA inside the human body.

Development of blood drug concentration/immunogenicity detection kit

In order to further support and facilitate your preclinical and clinical research of antibody drugs, we also offer our ELISA kit development services for immunogenicity (anti-drug antibody, ADA) or pharmacokinetic (PK) assays. Immunogenicity assays are used to quantify the level of immune response against the targeted therapeutic, while pharmacokinetic assays are used to elucidate the drug concentration and drug clearance within the body. Both assays are critical regulatory requirements for drug development, IND applications, clinical trials and post-marketing monitoring.

ACROBiosystems provides custom development services for both drug concentration and ADA detection kits for customers' diversified needs and applications while also providing key reagents for PK and immunogenicity analysis of biological drugs.

CAR-T
NamePeriodDeliveryPrice

Anti-idiotypic rabbit polyclonal antibody preparation

8-10 weeks

  • 1. Rabbit antiserum;

  • 2. Immunoaffinity chromatography purified polyclonal antibody (specific amount according to customer's demand), liquid/lyophilised product available,  buffer customised ;

  • 3. 1-3mg of synthetic antigen;

  • 4. Quality inspection report (COA);

  • 5. Complete experiment report.

One-on-one service of the project team
According to your needs
Issue a customized plan for you


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Anti-idiotypic mouse monoclonal antibody preparation

4-5 months

  • 1. positive cell lines;

  • 2. Ascites;

  • 3. purified antibodies (specific amount customised according to customer requirements);

  • 4. Quality inspection report (COA);

  • 5. Complete experiment report.

Development of a quantitative detection kit for blood drug concentration

5-7 weeks

  • 1. Methodology verification report;

  • 2. Instructions, COA;

  • 3. ELISA kit

ACRO Anti-idiotypic Antibody Products
MoleculeCat. No.AntigenNeutralizing ActivityApplication
Adalimu*abADB-Y19Anti-Adalimu*ab Antibodies (AY19)Neutralizing AntibodyADA assay; Neutralizing assay; Indirect ELISA
Bevacizu*abBEB-Y10Anti-Bevacizu*ab Antibodies (AY10) (MALS verified, recommended for PK/PD)Neutralizing AntibodyPK bridging ELISA; Neutralizing assay; Indirect ELISA
Bevacizu*abBEB-Y12Anti-Bevacizu*ab Antibodies (AY12) (recommended for neutralizing assay)Neutralizing AntibodyADA assay; Neutralizing assay; Indirect ELISA
Bevacizu*abBEB-Y9Anti-Bevacizu*ab Antibodies (AY9) (recommended for ADA assay)Neutralizing AntibodyADA assay; Neutralizing assay; Indirect ELISA
Bevacizu*abBEB-BY13Biotinylated Anti-Bevacizu*ab Antibodies (AY13) (recommended for PK/PD)Neutralizing AntibodyPK bridging ELISA;Neutralizing assay; Indirect ELISA
Cetuxi*abCEB-Y27Anti-Cetuxi*ab Antibodies (AY27) (recommended for ADA assay)Neutralizing AntibodyADA assay; Neutralizing assay; Indirect ELISA
Cetuxi*abCEB-Y31Anti-Cetuxi*ab Antibodies (AY31) (Non-Neutralizing)Non-Neutralizing AntibodyADA assay; Indirect ELISA
Cetuxi*abCEB-Y28Anti-Cetuxi*ab Antibodies (AY28)Neutralizing AntibodyADA assay; Neutralizing assay; Indirect ELISA
Cetuxi*abCEB-BY31Biotinylated Anti-Cetuxi*ab Antibodies (AY31) (recommended for PK/PD)Non-Neutralizing AntibodyPK bridging ELISA; Indirect ELISA
Rituxi*abRIB-Y36Anti-Rituxi*ab Antibodies (AY36) (recommended for ADA assay)Neutralizing AntibodyADA assay;Neutralizing assay; Indirect ELISA
Rituxi*abRIB-Y37Anti-Rituxi*ab Antibodies (AY37) (recommended for PK/PD)Neutralizing AntibodyPK bridging ELISA;Neutralizing assay;Indirect ELISA
Rituxi*abRIB-FY35cFITC-Labeled Anti-Rituxi*ab Antibodies, Mouse IgG1Neutralizing AntibodyADA assay;Neutralizing assay; Indirect ELISA
Trastuzu*abTRB-Y5bAnti-Trastuzu*ab Antibodies (AY5b) (recommended for PK/PD)Non-Neutralizing AntibodyNeutralizing Antibody
Trastuzu*abTRB-Y1bAnti-Trastuzu*ab Antibodies (AY1b) (recommended for PK/PD)Neutralizing AntibodyPK bridging ELISA; Neutralizing assay; Indirect ELISA
Development Process
Anti-idiotypic mouse monoclonal antibody preparation
Antigen

1. Antigen recheck (1 to 2 days)

SDS-PAGE purity recheck, UV concentration recheck

2. Antigen preparation (1 week, optional)

The customer provides full-length antibody drugs, we perform enzymatic digestion and purification to obtain F(ab) '2, and analyze the obtained F(ab)'2 for purity and activity, and provide a report on antigen preparation

Immunization and titer detection (8 weeks)

3. Immunization and titer detection (8 weeks)

Blood collection prior to immunization, 10 Balb/c mice immunization, titer detection, final blood collection, Immunization schedule and titer detection report provided; titer meets 1:72,9000

Immune titer detection
After immunizing Balb/c mice with F(ab ')2 for four times, the final titer reached  1:72,9000
After immunizing Balb/c mice with F(ab ')2 for four times, the final titer reached 1:72,9000
Cell fusion and screening (6-8 weeks)

4. Cell fusion and screening (6-8 weeks)

Spleen B cells from one or two mice with high titer are selected for electrofusion with SP2/0, positive clones screened by full-length antibody, isotype control antibody and human IgG are detected by indirect ELISA blocking ELISA is verifed cloning of competing and non-competing targets and performed 2-3 rounds of subcloning

ELISA screening clone
The positive clones were screened by indirect ELISA
The positive clones were screened by indirect ELISA
The antibody types were screened by blocking ELISA
The antibody types were screened by blocking ELISA
Antibody production and pairing(2-3 weeks)

5. Antibody production and pairing (2-3 weeks)

Hybridoma cell culture micro-purification and labeling, Sandwich paired validation, SDS-PAGE purity analysis, UV concentration determination, ELISA activity analysis and identification, PK assay development using sandwich method

Scale-up of antibody production and assay establishment (2-3 weeks)

6. Scale-up of antibody production and assay establishment (2-3 weeks)

Optimized paired antibodies are mass-produced, purified by SDS-PAGE analysis, UV concentration determination, ELISA activity analysis, PK Assay (standard curve, sensitivity, matrix interference validation, etc.) are established, and sensitivity meets the clinical PK assay requirements

7. Deliverables

- Serum before immunization, cellular supernatant
- Purified antibodies, cell lines
- Titer test report, fusion screening report, antibody identification report, COA, detailed project development report

Anti-idiotypic rabbit polyclonal antibody preparation
Antigen recheck

1. Antigen recheck (1 to 2 days)

SDS-PAGE purity recheck, UV concentration recheck

2. Antigen check (1 to 2 days)

The customer provides full-length antibody drugs, we perform enzymatic digestion and purification to obtain F(ab) '2, and analyze the obtained F(ab)'2 for purity and activity, and provide a report on antigen preparation

Enzymatic digestion detection
The activity of F(ab ')2 was almost no different from that of full-length antibody
The antibody binding activity tested after enzymatic digestion. The activity of F(ab ')2 was almost no different from that of full-length antibody, which remained above 85%.
Immunization and titer detection (8 weeks)

3. Immunization and titer detection (8 weeks)

Blood collection before immunization, immunization of New Zealand white rabbits, titer detection, final blood collection, Immunization schedule and titer detection report provided; titer meets 1:72,9000

Antibody purification (2-3 weeks)

4. Antibody purification (2-3 weeks)

- Antigen affinity chromatography
- Total human IgG purification
- Purification of isotype IgG (optional)

Antibody development and identification
Affinity-purified polyclonal antibodies specifically bind antibody drugs, cross-reactivity with isotype control <2%
Affinity-purified polyclonal antibodies specifically bind antibody drugs, cross-reactivity with isotype control<2%
The initial sensitivity of anti-idiotypic polyclonal antibody is usually less than 10ng/ml
The initial sensitivity of anti-idiotypic polyclonal antibody is usually less than 10ng/ml
Antibody identification and Assay establishment (1 week)

5. Antibody identification and Assay establishment (1 week)

- SDS-PAGE purity analysis, UV concentration determination
- Cross-reaction with isotype Human IgG is identified and analyzed by ELISA activity; cross-reactivity<2%
- ADA Assay establishment (sensitivity, MRD, etc.), sensitivity meets regulatory requirements.

Antibody development and identification
Antibody development and identification

6. Kit development

7. Deliverables

- Rabbit serum before immunization
- Purified antibodies, developed Kit
- Titer test report, antibody identification report, COA, detailed project development report

 
 
 
 
 

     
Case study
PK assay——Specific detection of antibody drug levels in vivo
Case study: bispecific antibody drug under PK assay with a sensitivity of 0.24ng/ml.
Immunization with full-length bispecific antibodies yielded blocking clones against Arm1 and Arm2.
Screened blocking clones against Arm 1
Clone1Clone2Clone3Clone4Clone5Clone6Clone7Clone8Clone9Clone10A0
OD0.1420.0630.1070.1430.2080.2640.230.1480.1490.221.601
blocking rate91.13%96.06%93.32%91.07%87.01%83.51%85.63%90.76%90.69%86.26%/
Clone11Clone12Clone13Clone14Clone15Clone16Clone17Clone18Clone19Clone20A0
OD0.2950.2280.1190.0980.0780.2290.5160.7090.9230.9651.601
blocking rate81.57%85.76%92.57%93.88%95.13%85.7%67.77%55.72%42.35%39.73%/
Screened blocking clones against Arm 2
Clone1Clone2Clone3Clone4Clone5Clone6Clone7Clone8Clone9Clone10A0
OD0.1090.2320.1780.1580.1460.2760.2450.1060.1650.1981.546
blocking rate92.95%84.99%88.49%89.78%90.56%82.15%84.15%93.14%89.33%87.19%/
Clone11Clone12Clone13Clone14Clone15Clone16Clone17A0
OD0.480.4950.3870.6580.0640.6511.0621.546
blocking rate68.95%67.98%74.97%57.44%95.86%57.89%31.31%/
The paired antibodies were selected from the blocking clones of Anti-Arm1 and Anti-Arm2 for sandwich method development as well as bispecific antibody drug PK detection, the standard curve is as follows:
PK assay——Specific detection of antibody drug levels in vivo

Immobilized anti-arm1 antibody can bind Bispecific antibody drugs, and then add Biotin- anti-arm2 antibody. Detection was performed using HRP-conjugated streptavidin with sensitivity of 0.24 ng/mL (Intact Assay).

Case study:ADC drug under PK assay with a sensitivity of 0.69ng/ml.
Naked F(ab)2 was used for immunization and the anti-idiotype antibody obtained after screening, and the ADC drug PK was detected together with Mouse Anti-MMAE Antibody, Mouse IgG1 (MALS verified) (Cat. No. MME-M5252). The standard curve is as follows:
PK assay——Specific detection of antibody drug levels in vivo

Immobilized Anti- drug Antibody, Mouse IgG1 can bind ADC antibody drugs, and then add Biotin- Mouse Anti-MMAE Antibody, Mouse IgG1 (MALS verified). Detection was performed using HRP-conjugated streptavidin with sensitivity of 0.69 ng/mL (Intact Assay).

Comparison of different methods for detecting antibody drug levels in vivo
Testing methodCoatedSampleTesting
Antigen capture ELISACD20Antibodies to be testedGoat anti-human IgG
Anti-idiotypic capture ELISAAnti-Ritux*mab AntibodiesAntibodies to be testedGoat anti-human IgG
Bridging ELISA by anti-idiotypic antibodiesAnti-Ritux*mab AntibodiesAntibodies to be testedBiotinylated Anti-Ritux*mab Antibodies
Figure1. Detection of Ritux*mab by antigen-capture ELISA (0.1% serum).
Figure2. Detection of Ritux*mab by anti-idiotypic capture ELISA (0.1% serum).
Figure3. Detection of Ritux*mab by anti-idiotypic bridging ELISA (10% serum).
Testing methodLinear range
(µg/mL)
Sensitivity
(µg/mL)
AdvantageDisadvantage
Antigen-capture ELISASimple method and good versatilityHigh background, no activity
Anti-idiotypic capture ELISA0.156-100.156Solve the difficulty in obtaining CD20, simple methodHigh background, only suitable for Rituxa biosimilar
Bridging ELISA by anti-idiotypic antibodies0.012-0.780.012Solve the difficulty in obtaining CD20, good sensitivity and low backgroundOnly applicable to Rituxan biosimilar
ADA assay—Immunogenicity analysis
The case of rabbit polyclonal antibody
Case study: Antibody cross-reaction verification

New Zealand white rabbits were immunized with full-length monoclonal antibodies. The antiserum was affinity purified for polyclonal antibodies specific to the drug. Cross reactivity to subtype control was less than 2%.

Case study:Antidrug antibody potency test for peptides

New Zealand white rabbits were immunized with polypeptide-KLH conjugates. Antiserum was purified by affinity chromatography until antibody titer was more than 1:712,000.

The case of mouse monoclonal antibody

Anti-Adalimumab Antibodies bridging ELISA for Anti-Drug Antibody (ADA) assay development. Immobilized adalimumab at 1 µg/ml, add increasing concentrations of Anti-Adalimumab Antibodies (Cat. No. ADB-Y19, 10% human serum) and then add biotinylated adalimumab at 5 µg/ml. Detection was performed using HRP-conjugated streptavidin with a sensitivity of 0.6 ng/mL.

Anti-Rituximab Antibodies bridging ELISA for Anti-Drug Antibody (ADA) assay development. Immobilized rituximab at 1 µg/ml, added increasing concentrations of Anti-Adalimumab Antibodies (Cat. No. RIB-Y36, 10% human serum) and then added biotinylated rituximab at 2 µg/ml. Detection was performed using HRP-conjugated streptavidin with a sensitivity of 9.7 ng/mL.

Inquiry and order

  • ACROBiosystems scientist will respond within 24 hours of submission.

  • Call us at: +1 800-810-0816 or email at services@acrobiosystems.com for consultation.

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