Name: Human IgG4-S228P Isotype Control Antibody
Brand: Syd Labs
SKU: PA007128
Price: 150.00 USD
Availability: InStock
Rating: 5 (10 reviews)

Catalog No.

Product Name

Size

List Price (US$)

Quantity

PA007128

In Vivo Grade Recombinant Human IgG4-S228P Isotype Control Antibody

1 mg

150.00

PA007128

In Vivo Grade Recombinant Human IgG4-S228P Isotype Control Antibody

5 mg

300.00

PA007128

In Vivo Grade Recombinant Human IgG4-S228P Isotype Control Antibody

25 mg

800.00

PA007128.05

In Vivo Grade Recombinant Human IgG4-S228P Isotype Control Antibody, Endotoxin 0.05 EU/mg

5 mg

450.00

PA007128.05

In Vivo Grade Recombinant Human IgG4-S228P Isotype Control Antibody, Endotoxin 0.05 EU/mg

25 mg

1200.00

PA007131

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

1 mg

150.00

PA007131

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

5 mg

300.00

PA007131

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

25 mg

800.00

PA007131.05

In Vivo Grade Recombinant Human IgG4-S228P L235E P329G (SPLEPG) Isotype Control Antibody, Endotoxin 0.05 EU/mg

5 mg

450.00

PA007131.05

In Vivo Grade Recombinant Human IgG4-S228P L235E P329G (SPLEPG) Isotype Control Antibody, Endotoxin 0.05 EU/mg

25 mg

1200.00

Description

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:

A Phase 1 Single-Ascending-Dose Trial in Healthy Volunteers to Evaluate the Safety, Tolerability, Pharmacokinetics, and Immunogenicity of Intravenous PNT001, a Novel Mid-domain Tau Antibody Targeting cis-pT231 Tau
Luca, W., et al. J Prev Alzheimers Dis. 2024;11(1):227-235. doi: 10.14283/jpad.2024.25. PMID: 38374743
PNT001 is a humanized full-length IgG4 S228P monoclonal antibody that binds the cis conformation of the phosphorylated Thr231-Pro232 motif in human full-length (2N4R) tau (cis-pT231 tau) with high selectivity and affinity. It binds selectively to cis-pT231 tau in human tauopathy brain sections, inhibits aggregation of tau, and has shown efficacy in preclinical models of tauopathy. Doses of 900-4,000 mg produced concentrations in the cerebrospinal fluid exceeding the binding affinity constant of PNT001 for cis-pT231 tau (45 ng/mL), indicating that concentrations sufficient for target engagement can be obtained in the cerebrospinal fluid within the tested dose range. The primary objective of this study was to evaluate the safety and tolerability of single-escalating i.v. doses of PNT001 in healthy volunteers. The results of this first-in-human study of PNT001 showed a tolerable safety profile of PNT001 after a single dose in healthy volunteers over a dose range of 33–4,000 mg.
Tags: activity of Human IgG4-S228P Isotype Control; activity of Human IgG4-S228P Isotype Control mAb

Fc-Fc interactions and immune inhibitory effects of IgG4: implications for anti-PD-1 immunotherapies
Zhang, W., et al. J Immunother Cancer. 2024 Jun 17;12(6):e009034. doi: 10.1136/jitc-2024-009034. PMID: 38925680
The majority of anti-programmed cell-death 1 (PD-1) monoclonal antibodies (mAbs) use S228P mutation IgG4 as the structural basis to avoid the activation of immune cells or complement. Wildtype IgG1 and IgG4, anti-PD-1 mAb nivolumab (IgG4 S228P), penpulimab (Fc-null IgG1), and tislelizumab (Fc-null IgG4 S228P-R409K) were assessed for their binding reactions to the immobilized IgG proteins and quantitative kinetic data were obtained. IgG4 and IgG4 S228P anti-programmed cell-death 1 monoclonal antibody (mAb) bind to immobilized IgG through Fc–Fc interactions. The binding of IgG4wt and IgG4 S228P anti-programmed cell-death 1 monoclonal antibody to IgG1 antibodies immobilized on cancer cells. The Fc-functional IgG4 S228P anti-PD-1 mAb retains Fc–Fc interactions and Fc:FcR interaction like wildtype IgG4.
Tags: human IgG1 recombinant isotype control; Human IgG4-S228P Isotype Control antibody for cancer research

A new approach to produce IgG4-like bispecific antibodies
Zhao, C., et al. Sci Rep. 2021 Sep 10;11(1):18150. doi: 10.1038/s41598-021-97393-2. PMID: 34545109
We selected IgG 4 S228P as the development target (PDB: 5DK3) and analyzed the overlay crystal structures of CH1/CL and TCR Cα/Cβ (PDB: 3ARB) (Fig. 1A). To produce a bispecific antibody targeting both PD-1 and LAG-3, we selected IgG 4 S228P 32 to make the construct. As shown in Table 1, the binding kinetics of the antibody to different Fcγ receptors are indistinguishable from the two parental antibodies and an isotype control of human IgG 4 S228P. The thermostability was also examined with Uncle biologics stability screening platform (Unchained Labs). As shown in Supplementary Fig. S5C, the melting temperature (Tm1) is 62.37 °C, lower than the control IgG 4 S228P whose Tm1 is 69.00 °C, indicating that PD-1 × LAG3 TiMab is relatively stable.
Tags: Human IgG4-S228P Isotype Control antibody for mouse tumor model; Human IgG4-S228P Isotype Control antibody in vivo

Impact of IgG subclass on molecular properties of monoclonal antibodies
Tang, Y., et al. MAbs. 2021 Dec;13(1):1993768. doi: 10.1080/19420862.2021.1993768. PMID: 34763607
To further establish mechanistic understandings, we conducted a systematic study by grafting three independent variable regions onto human IgG1, an IgG1 variant, IgG2, and an IgG4 variant constant domains and evaluating the impact of subclass and variable regions on their molecular properties. Surface charge analysis at pH 6 (3a and 3b) and hydrophobic patch analysis (3c) for the antibodies with variable region of Fv A, B, and C (3a) in the subclass structure of IgG1, IgG1EN, IgG2, and IgG4PAA by homology modeling. Our data indicate that IgG subclass plays a significant role on molecular properties, either through direct effects or via the interplay with the variable region, the IgG1 mAbs tend to have higher solubility than either IgG2 or IgG4 mAbs in a common pH 6 buffer matrix, and solution behavior relies heavily on the charge status of the antibody at the desirable pH. Structural and computational analysis revealed specific molecular features that potentially account for the differential behavior of the IgG subclasses observed experimentally.
Tags: Human IgG4-S228P Isotype Control for cancer research; Human IgG4-S228P Isotype Control for mouse tumor model

Comprehensive Analysis of the Therapeutic IgG4 Antibody Pembrolizumab: Hinge Modification Blocks Half Molecule Exchange In Vitro and In Vivo
Yang, X., et al. J Pharm Sci. 2015 Nov;104(11):3511-20. doi: 10.1002/jps.24620. PMID: 26308749
However, human IgG4 can undergo Fab arm (half molecule) exchange with other IgG4 molecules in vivo. The hinge modification by a point mutation (S228P) prevents half molecule exchange of IgG4. Both in vitro and in vivo results indicate that the hinge modification in pembrolizumab prevented half molecule exchange, whereas the unmodified counterpart anti-PD1 wt showed active exchange activity with other IgG4 antibodies or self-exchange activity with its own molecules. Our work, as an example expected for meeting regulatory requirements, contributes to establish without ambiguity that hinge-modified IgG4 antibodies are suitable for biotherapeutic applications.
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.

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

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Human IgG4-S228P Isotype Control Antibody

hIgG4 Isotype Control, Human IgG4 Isotype Control, hIgG4 S228P, hIgG4 SPLEPG Mutant

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References of Human IgG4-S228P Isotype Control Antibody: