Antibody Affinity Purification Resin Solutions: Principles, Products, and Applications

08 Jan 2026 by AntBio

I. Background Introduction

Antibody affinity purification is a core technology in life science research and biopharmaceutical production, enabling high-purity and high-yield isolation of antibodies or antigen-antibody complexes. The key to this technology lies in affinity ligands immobilized on resin matrices, which specifically bind to antibodies. Common affinity ligands include Protein A, Protein G, Protein A/G, and Protein L, each with unique binding characteristics and application scopes:

• Protein A: A surface protein from Staphylococcus aureus, composed of five homologous Ig-binding domains folded into triple-helical bundles.

• Protein G: A protein from the cell wall of group G and C streptococci.

• Protein A/G: A recombinant fusion protein of Protein A and Protein G, incorporating 5 Fc-binding domains from Protein A and 3 Fc-binding domains from Protein G.

• Protein L: Derived from Peptostreptococcus magnus, it binds to immunoglobulins through interactions with the light chain (κ chain).

Figure 1 Binding Regions Between Antibodies and Antibody Affinity Ligands

Table 1 Analysis of Antibody Affinity Differences Among Protein A, G, A/G, and L

Ligand Type	Advantages	Disadvantages	Applications
Protein A	Binding sites are located in the Fc region between the CH2 and CH3 domains, as well as in the heavy chain within the Fab region of the human VH3 family.	-	Purification, immobilization, or detection of immunoglobulins.
Protein G	Can specifically bind to the Fc segment of immunoglobulin IgG molecules without affecting the ability of the Fab segment to bind to antigen molecules.	-	Isolation and purification of IgG from serum, purification of monoclonal antibodies, and separation of antigen-antibody complexes.
Protein A/G	Can bind to all human IgG subtypes, as well as IgA, IgE, and IgM. Protein A/G reduces pH dependence, allowing binding at pH 5-8.	Cannot bind to mouse IgA, IgM, or serum albumin.	Extraction and detection of mouse IgG monoclonal antibodies.
Protein L	Only binds to the κ light chain of antibodies without affecting the antigen-binding site of the antibody. Can bind to a wider range of antibody classes, such as human IgG, IgM, IgA, IgE, IgD, as well as antibody fragments containing κ light chains (human types 1, 3, 4; mouse type 1), such as Fab and scFv.	Does not bind to immunoglobulins from cattle, goats, or sheep.	Purification of monoclonal antibodies from culture media supplemented with BSA or FCS, and widely used in the purification and separation of genetically engineered antibodies.

Table 2 Affinity Between Antibody Affinity Ligands and Antibodies of Different Origins and Subtypes

Species	Subtype	Protein A	Protein G	Protein A/G	Protein L
Human	IgG1	+++	+++	+++	+++
	IgG2	+++	+++	+++	+++
	IgG3	+	+++	+++	+++
	IgG4	+++	+++	+++	+++
	IgM	+	-	+	+++
	IgD	-	-	-	+++
	IgA	+	-	+	+++
	Fab	+	+	+	+++
Mouse	IgG1	+	++	++	+++
	IgG2a	+++	+++	+++	+++
	IgG2b	+++	+++	+++	+++
	IgG3	+++	+++	+++	+++
	IgM	-	-	-	+++
Rat	IgG1	+	++	++	+++
	IgG2a	-	+++	+++	+++
	IgG2b	-	+	+	+++
	IgG2c	+++	+++	+++	+++
Bovine	IgG1	+	+++	+++	-
Bovine	IgG2	+++	+++	+++	-
Goat	IgG1	+	+++	+++	-
Goat	IgG2	+++	+++	+++	-
Horse	IgG(ab)	+	-	+	NA
	IgG(c)	+	-	+	NA
	IgG(T)	-	+++	+++	NA
Rabbit	IgG	+++	+++	+++	+
Pig	IgG	+++	+	+++	+++
Dog	IgG	+++	+	+++	NA
Chicken	IgY	-	-	-	-

Note: - indicates no binding; + indicates moderate binding; +++ indicates strong binding; NA indicates not applicable or data unavailable.

II. Parameters of Antibody Affinity Purification Resins

Category	Recombinant Protein A Sepharose FF	Recombinant Protein G Sepharose FF	High Alkali-Resistant Protein A Sepharose FF	Protein A/G Sepharose FF	Recombinant Protein L Sepharose FF
Matrix	4% cross-linked agarose	4% cross-linked agarose	4% cross-linked agarose	4% cross-linked agarose	4% cross-linked agarose
Ligand	Recombinant Staphylococcus aureus Protein A, ≥6mg/mL	Recombinant Streptococcus Protein G, ≥6mg/mL, single-point conjugation	Recombinant alkali-resistant Protein A, ≥6mg/mL	Protein A/G fusion protein, ≥6mg/mL	Recombinant Peptostreptococcus magnus Protein L, ≥6mg/mL
Particle Size Range	45-165μm	45-165μm	45-165μm	45-165μm	45-165μm
Average Particle Size	~90μm	~90μm	~90μm	~90μm	~90μm
Dynamic Binding Capacity	≥40mg h-IgG/mL	≥40mg h-IgG/mL	≥45mg h-IgG/mL	≥40mg h-IgG/mL	≥40mg κ light chain h-IgG/mL
Recommended Operating Flow Rate	60-300cm/h	60-300cm/h	60-300cm/h	60-150cm/h	60-300cm/h
Maximum Flow Rate and Pressure	＞900cm/h	＞900cm/h	＞900cm/h	＞900cm/h	＞900cm/h
Operating pH	3-10 (recommended operating pH), 3-12 (short-term stability)	3-9 (recommended operating pH), 2-10 (short-term stability)	3-10 (recommended operating pH), 3-12 (short-term stability)	3-10 (recommended operating pH), 3-12 (short-term stability)	2-9 (recommended operating pH), 15mM NaOH (CIP)
Chemical Stability	Stable in the following solutions: common aqueous buffers, 10-100mmol/L NaOH, 6mol/L guanidine hydrochloride, 70% ethanol.	Stable in the following solutions: common aqueous buffers, 6mol/L guanidine hydrochloride, 70% ethanol.	Stable in the following solutions: common aqueous buffers, 0.1-0.5mol/L sodium hydroxide, 6mol/L guanidine hydrochloride, 70% ethanol, etc.	Stable in the following solutions: common aqueous buffers, 0.1-0.5mol/L sodium hydroxide, 6mol/L guanidine hydrochloride, 70% ethanol, etc.	Stable in the following solutions: common aqueous buffers, 6mol/L guanidine hydrochloride, 70% ethanol.

Notes:

a: More than 90% by volume of microspheres are within this particle size range;

b: DBC10% test conditions: 10% breakthrough, 6min residence time;

c: The recommended flow rate refers to the flow rate that can meet the usage conditions of 2-6min residence time under most column heights, not that it can only work within this flow rate range;

d: Maximum test flow rate under 10cm column height.

III. Application Data

Sample: Bovine serum, approximately 20mg IgG/mL resin relative to the resin;

Loading and Binding Buffer: 20mM PB, 150mM NaCl, pH7.4, 10CV;

Elution Buffer: 100mM Glycine, pH3.0, 14CV;

Cleaning in Place (CIP): 100mM NaOH, 10CV

IV. Antibody Affinity Purification Resin Product Information of ANT BIO PTE. LTD.

Product Code	Product Name	Specification
abs91001-F	Recombinant Protein A Sepharose FF	10mL/100mL
abs91001-G	Recombinant Protein A Sepharose FF (Gravity Prepacked Column)	1mL/5mL
abs91001-M	Recombinant Protein A Sepharose FF (Medium Pressure Prepacked Column)	1mL/5mL
abs91002-F	Recombinant Protein G Sepharose FF	10mL/100mL
abs91002-G	Recombinant Protein G Sepharose FF (Gravity Prepacked Column)	1mL/5mL
abs91002-M	Recombinant Protein G Sepharose FF (Medium Pressure Prepacked Column)	1mL/5mL
abs91003-F	High Alkali-Resistant Protein A Sepharose FF	10mL/100mL
abs91003-M	High Alkali-Resistant Protein A Sepharose FF (Medium Pressure Prepacked Column)	1mL/5mL
abs91003-G	High Alkali-Resistant Protein A Sepharose FF (Gravity Prepacked Column)	1mL/5mL
abs91004-F	Protein A/G Sepharose FF	10mL/100mL
abs91004-M	Protein A/G Sepharose FF (Medium Pressure Prepacked Column)	1mL/5mL
abs91004-G	Protein A/G Sepharose FF (Gravity Prepacked Column)	1mL/5mL
abs91005-F	Easy Elution Protein G Sepharose FF	10mL/100mL
abs91005-M	Easy Elution Protein G Sepharose FF (Medium Pressure Prepacked Column)	1mL/5mL
abs91005-G	Easy Elution Protein G Sepharose FF (Gravity Prepacked Column)	1mL/5mL
abs91006-F	Recombinant Protein L Sepharose FF	10mL/100mL
abs91006-M	Recombinant Protein L Sepharose FF (Medium Pressure Prepacked Column)	1mL/5mL
abs91006-G	Recombinant Protein L Sepharose FF (Gravity Prepacked Column)	1mL/5mL

Brand Mission

ANT BIO PTE. LTD. is dedicated to advancing life science research and biopharmaceutical development by providing high-quality, reliable antibody affinity purification resin solutions. We recognize the critical role of antibody purification in scientific research and industrial production, and the urgent need for high-performance, stable, and versatile purification tools. Through our specialized sub-brands (Absin, Starter, UA), we have developed a comprehensive product portfolio of antibody affinity purification resins, covering Protein A, Protein G, Protein A/G, and Protein L series, to meet the diverse needs of antibody purification from different species and subtypes.

Our team adheres to stringent quality control standards throughout the product development and production process, ensuring the high ligand density, excellent dynamic binding capacity, and chemical stability of each resin product. We are committed to providing professional technical support and customer-centric services, helping researchers and production personnel overcome challenges such as low purification efficiency, poor product stability, and difficult cleaning, and accelerating the pace of scientific research breakthroughs and biopharmaceutical production. ANT BIO PTE. LTD. strives to be a trusted partner for scientists and biopharmaceutical companies worldwide, contributing to the advancement of life science research and the development of innovative therapeutic drugs.

Disclaimer

This article is AI-compiled and interpreted based on the original work related to antibody affinity purification resin research. All intellectual property (e.g., images, data, experimental protocols) of the original publication shall belong to the journal and the research team. For any infringement, please contact us promptly and we will take immediate action.

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At ANTBIO, we are committed to advancing life science research through high-quality, reliable reagents and comprehensive solutions. Our specialized sub-brands (Absin, Starter, UA) cover a full spectrum of research needs, from antibody affinity purification resins and general reagents to kits and antibodies. With a focus on innovation, quality, and customer-centricity, we strive to be your trusted partner in unlocking scientific mysteries and driving medical progress. Explore our product portfolio today and elevate your research to new heights.