Human IgG4-S228P Isotype Control Antibody

The ideal recombinant human IgG4 isotype control antibody (hIgG4 S228P kappa isotype control) has low or no specific binding to any human sample, and humanized or completely human variable regions in addition to the human IgG4 S228P kappa constant regions. The human IgG4 kappa isotype control and mutants can be used in the cell-based assay or animal model as the antibody negative control when evaluating fully human, humanized, or chimeric antibodies.

The hIgG4 isotype control mutants were generated because the corresponding mutations had been introduced to some human IgG4 therapeutic antibody candidates. For example, S228P, an engineered mutation in the core hinge region, is used to prevent Fab-arm exchange in vivo of human IgG4 molecules; P329G substitution combined with S228P and L235E was shown to decrease effector functions by disrupting the Fc/Fcγ receptor interface. When clients test their engineered antibody candidates, the silent Fc effctor function human IgG4 isotype controls are useful for in vivo use.

Welcome to contact us and ask for a quote for the engineered human IgG4 isotype control mutants, hIgG4 isotype control and Fc silenced mutants with Avi-, His-, and DYKDDDDK-tags, and biotinylated hIgG4 isotype control and silent Fc mutants. A variety of conjugates (such as dyes and fluorophores) with the human IgG4 isotype control and silent Fc effctor function mutants are available.

PA007128: In Vivo Grade Recombinant Human IgG4-S228P Isotype Control Antibody (hIgG4 Isotype Control)

The recombinant human IgG4-S228P kappa isotype control antibody (hIgG4 S228P isotype control) was produced in CHO cells.
Immunogen: Trinitrophenol (TNP).
Clone: 1R12.
Isotype: human IgG4, kappa.
Applications: an isotype-matched negative control for human IgG4 kappa antibodies used in ELISA, Western Blot (WB), Flow Cytometry (Flow), Immunoprecipitation (IP), Immunohistochemistry (Paraffin) (IHC (P)), Immunohistochemistry (Frozen) (IHC (F)), and in vivo animal model research.
Formulation: 0.2 μM filtered solution (1 mg/ml) of 1x PBS.
Purity: >95% by SDS-PAGE under reducing conditions.
Endotoxin Level: Less than 1 EU/mg of protein as determined by LAL method. Endotoxin ≤0.05 EU/mg for PA007128.05, in vivo grade recombinant human IgG4-S228P isotype control antibody, premium grade (endotoxin ≤0.05 EU/mg).

PA007131: In Vivo Grade Recombinant Human IgG4-S228P L235E P329G Isotype Control Antibody (hIgG4 SPLEPG Isotype Control Mutant)

The recombinant human IgG4-S228P L235E P329G isotype control antibody (hIgG4 SPLEPG isotype control mutant, Fc silenced) was produced in mammalian cells.
Immunogen: N/A.
Clone: 4F17.
Isotype: human IgG4, kappa.
Applications: an isotype-matched negative control for human IgG4 SPLEPG kappa antibodies used in ELISA, Western Blot (WB), Flow Cytometry (Flow), Immunoprecipitation (IP), Immunohistochemistry (Paraffin) (IHC (P)), Immunohistochemistry (Frozen) (IHC (F)), and in vivo animal model research.
Formulation: 0.2 μM filtered solution of 1x PBS.
Purity: >95% by SDS-PAGE under reducing conditions.
Endotoxin Level: Less than 1 EU/mg of protein as determined by LAL method.

Shipping: The recombinant hIgG4 kappa isotype control and mutants are shipped with ice pack. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
1 month from date of receipt, 2 to 8°C as supplied.
3 months from date of receipt, -20°C to -70°C as supplied.

Background

Isotype control antibodies are used as negative controls of primary antibodies to check presence of non-specific binding to FC receptors on the cell surface, other cellular proteins, carbohydrates, and lipids. Thus, the isotype control antibody normally matches the host species and isotype, including light chains (such as human IgG4 S228P kappa antibody), concentration, and conjugation format of the experimental primary antibody. The ideal human isotype control antibody has low or no specific binding to any human sample, and humanized or completely human variable regions in addition to the human IgG constant regions.

Isotype control antibodies are frequently used in IHC and Flow cytometry experiments, and in vivo animal study.

References of Human IgG4-S228P Isotype Control Antibody:

Modulation of urelumab glycosylation separates immune stimulatory activity from organ toxicity.
Reitinger, C., et al. Front Immunol. 2022 Sep 29;13:970290. doi: 10.3389/fimmu.2022.970290. PMID: 36248847
Thus, choosing IgG subclasses, such as human IgG4, with a low ability to interact with FcγRs has been identified as a potential strategy to limit FcγR or complement pathway dependent side-effects. ...However, even immunomodulatory antibodies on the human IgG4 background may interact with cellular FcγRs and show dose limiting toxicities. ...Thus, glycoengineering of human IgG4 antibodies may be a possible approach to limit collateral damage by immunomodulatory antibodies and allow for a greater therapeutic window of opportunity. ...However, several studies have shown that human IgG4 antibodies may productively interact with human FcγRs in vitro and in vivo, suggesting that human IgG4 Fc-domains are not inert and may contribute to wanted and unwanted effects of immunomodulatory antibodies in vivo. ...The human IgG4 CD137-specific antibody urelumab has shown promising results in pre-clinical and clinical settings.
Tags: activity of Human IgG4-S228P Isotype Control; activity of Human IgG4-S228P Isotype Control mAb

Targeting PD-L1 in solid cancer with myeloid cells expressing a CAR-like immune receptor.
Myers Chen, K., et al. Front Immunol. 2024 Apr 25;15:1380065. doi: 10.3389/fimmu.2024.1380065. PMID: 38726005
In some conditions, macrophages were pre-treated with 2μM cytochalasin D, 10μg/mL anti-PD1 antibody or 10μg/mL human IgG4 isotype control antibody prior to co-culture with target cells. ...To evaluate CARIR-mediated killing activity on PD-L1+ target cells, non-modified or CARIR engineered THP-1 cells (2.5 × 104) were co-cultured with either RM-1hPD-L1 or RM-1 target cells (5 × 103) in the presence of 5ng/ml PMA in 96-well round-bottom culture plate for 3 days. ...Solid cancers Myeloid cells are prevalent in solid cancers, but they frequently exhibit an anti-inflammatory pro-tumor phenotype that contribute to the immunosuppressive tumor microenvironment (TME), which hinders the effectiveness of cancer immunotherapies. ...One such engineering approach is to target the checkpoint molecule PD-L1, which is often upregulated by solid cancers to evade immune responses. ...Here we devised an adoptive cell therapy strategy based on myeloid cells expressing a Chimeric Antigen Receptor (CAR)-like immune receptor (CARIR).
Tags: human IgG1 recombinant isotype control; Human IgG4-S228P Isotype Control antibody for cancer research

R409K mutation prevents acid-induced aggregation of human IgG4.
Namisaki, H., et al. PLoS One. 2020 Mar;15(3):e0229027. doi: 10.1371/journal.pone.0229027. PMID: 32182240
Currently, therapeutic IgG4 antibodies, such as nivolumab, are adapted from a human IgG4 variant containing a substitution, S228P, to prevent formation of half-antibodies. ...No previous study has comprehensively analyzed the specific amino acid residues in human IgG4 responsible for acid-induced aggregation. ...We also used amino acid substitutions to establish a stable IgG4 variant without altering the unique biological characteristics of human IgG4-format antibodies used in therapeutics. ...To identify the domain responsible for aggregation in human IgG4, we performed domain-subclass conversion of the CH regions. ...Human IgG4 antibodies are suitable for use in the potential low-effector-function format of therapeutic antibodies.
Tags: in vivo study of Human IgG4-S228P Isotype Control antibody; bulk Human IgG4-S228P Isotype Control

For more references about Human IgG4-S228P Isotype Control Antibody please contact our scientific support team with message@sydlabs.com.

Other in vivo grade Recombinant IgG Isotype Control Antibodies and Mutants:
In vivo Grade Recombinant Human IgG1 Isotype Control Antibody and Mutants
In vivo Grade Recombinant Human IgG2 Isotype Control Antibody
In vivo Grade Recombinant Human IgG3 Isotype Control Antibody
In vivo Grade Recombinant Human IgG4-S228P Isotype Control Antibody and Mutants
In vivo Grade Recombinant Mouse IgG1 Isotype Control Antibody and Mutants
In vivo Grade Recombinant Mouse IgG2a Isotype Control Antibody and Mutants
In vivo Grade Recombinant Mouse IgG2b Isotype Control Antibody and Mutants
In vivo Grade Recombinant Mouse IgG2c Isotype Control Antibody and Mutants
In vivo Grade Recombinant Mouse IgG3 Isotype Control Antibody
In vivo Grade Recombinant Rat IgG1 Isotype Control Antibody
In vivo Grade Recombinant Rat IgG2a Isotype Control Antibody
In vivo Grade Recombinant Rat IgG2b Isotype Control Antibody
In vivo Grade Recombinant Rat IgG2c Isotype Control Antibody
In vivo Grade Recombinant Hamster IgG1 Isotype Control Antibody
In vivo Grade Recombinant Hamster IgG2 Isotype Control Antibody

In vivo Grade Recombinant IgG Fc Proteins:
In vivo Grade Recombinant Human IgG1 Fc Protein (hIgG1)
In vivo Grade Recombinant Human IgG2 Fc Protein (hIgG2)
In vivo Grade Recombinant Human IgG4 Fc Protein (hIgG4)
In vivo Grade Recombinant Mouse IgG1 Fc Protein (mIgG1)
In vivo Grade Recombinant Mouse IgG2a Fc Protein (mIgG2a)
In vivo Grade Recombinant Mouse IgG2b Fc Protein (mIgG2b)
In vivo Grade Recombinant Rat IgG2a Fc Protein (rtIgG2a)
In vivo Grade Recombinant Rat IgG2b Fc Protein (rtIgG2b)
In vivo Grade Recombinant Llama IgG2b Fc Protein (lIgG2b)
In vivo Grade Recombinant Rabbit IgG Fc Protein (rIgG)

Fc ELISA Kits and Reagents:
Human Fc ELISA Kit
Mouse Fc ELISA Kit
Human Fc ELISA Reagent Kit
Mouse Fc ELISA Reagent Kit

Protein A, Resin, Columns, and ELISA Kits:
Recombinant Protein A
Protein A Affinity Resin
Pre-packed Protein A Column
Protein A ELISA Kit