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From rabbit antibody repertoires to rabbit monoclonal antibodies

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ABSTRACT

In this review, we explain why and how rabbit monoclonal antibodies have become outstanding reagents for laboratory research and increasingly for diagnostic and therapeutic applications. Starting with the unique ontogeny of rabbit B cells that affords highly distinctive antibody repertoires rich in in vivo pruned binders of high diversity, affinity and specificity, we describe the generation of rabbit monoclonal antibodies by hybridoma technology, phage display and alternative methods, along with an account of successful humanization strategies.

No MeSH data available.


Related in: MedlinePlus

Schematic drawing of natural rabbit antibodies in IgG format. The ~150-kDa rabbit IgG molecule contains two identical κ (white) or λ (light gray) light chains paired with two identical heavy chains (dark gray). The light chain consists of an N-terminal variable domain (VL), shown with its three CDRs, followed by one constant domain (CL). The heavy chain consists of an N-terminal variable domain (VH), also shown with its three CDRs, followed by three constant domains (CH1, CH2 and CH3). CH1 and CH2 are linked through a flexible hinge region that has the amino-acid sequence APSTCSKPTCP (or APSTCSKPMCP in an allotypic variant) and anchors three disulfide bridges (orange) of the IgG molecule, one for each of the two light- and heavy-chain pairs, and one for the heavy-chain pair. Notably, rabbits have two κ light chains, K1 and K2. The more frequent κ light chain, K1, contains an additional disulfide bridge that links VL and CL. Rabbits of the commonly used New Zealand White strain have ~90% IgG-κ (K1), ~10% IgG-κ (K2) and <1% IgG-λ antibodies. CDR, complementarity-determining region.
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fig2: Schematic drawing of natural rabbit antibodies in IgG format. The ~150-kDa rabbit IgG molecule contains two identical κ (white) or λ (light gray) light chains paired with two identical heavy chains (dark gray). The light chain consists of an N-terminal variable domain (VL), shown with its three CDRs, followed by one constant domain (CL). The heavy chain consists of an N-terminal variable domain (VH), also shown with its three CDRs, followed by three constant domains (CH1, CH2 and CH3). CH1 and CH2 are linked through a flexible hinge region that has the amino-acid sequence APSTCSKPTCP (or APSTCSKPMCP in an allotypic variant) and anchors three disulfide bridges (orange) of the IgG molecule, one for each of the two light- and heavy-chain pairs, and one for the heavy-chain pair. Notably, rabbits have two κ light chains, K1 and K2. The more frequent κ light chain, K1, contains an additional disulfide bridge that links VL and CL. Rabbits of the commonly used New Zealand White strain have ~90% IgG-κ (K1), ~10% IgG-κ (K2) and <1% IgG-λ antibodies. CDR, complementarity-determining region.

Mentions: European rabbits (Oryctolagus cuniculus; Figure 1) have played an important role as animal models in immunology for many decades.1, 2 Today, rabbits are still a major source for a wide variety of monoclonal antibodies (mAbs) and polyclonal antibodies (pAbs) with broad utility. pAbs can be described as a set of different antibodies generated in response to a specific pathogen or antigen, generally targeting different epitopes. mAbs, on the other hand, contain a defined antigen-binding site (paratope) that typically binds with high affinity and specificity to only one epitope. From a pharmaceutical point of view, mAbs provide a molecularly defined and reproducible product, whereas pAbs are traditionally an imprecise mixture of different antibodies.3 As is the case for mouse and human mAbs, IgG is the most common isotype of rabbit mAbs (Figure 2).


From rabbit antibody repertoires to rabbit monoclonal antibodies
Schematic drawing of natural rabbit antibodies in IgG format. The ~150-kDa rabbit IgG molecule contains two identical κ (white) or λ (light gray) light chains paired with two identical heavy chains (dark gray). The light chain consists of an N-terminal variable domain (VL), shown with its three CDRs, followed by one constant domain (CL). The heavy chain consists of an N-terminal variable domain (VH), also shown with its three CDRs, followed by three constant domains (CH1, CH2 and CH3). CH1 and CH2 are linked through a flexible hinge region that has the amino-acid sequence APSTCSKPTCP (or APSTCSKPMCP in an allotypic variant) and anchors three disulfide bridges (orange) of the IgG molecule, one for each of the two light- and heavy-chain pairs, and one for the heavy-chain pair. Notably, rabbits have two κ light chains, K1 and K2. The more frequent κ light chain, K1, contains an additional disulfide bridge that links VL and CL. Rabbits of the commonly used New Zealand White strain have ~90% IgG-κ (K1), ~10% IgG-κ (K2) and <1% IgG-λ antibodies. CDR, complementarity-determining region.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5382564&req=5

fig2: Schematic drawing of natural rabbit antibodies in IgG format. The ~150-kDa rabbit IgG molecule contains two identical κ (white) or λ (light gray) light chains paired with two identical heavy chains (dark gray). The light chain consists of an N-terminal variable domain (VL), shown with its three CDRs, followed by one constant domain (CL). The heavy chain consists of an N-terminal variable domain (VH), also shown with its three CDRs, followed by three constant domains (CH1, CH2 and CH3). CH1 and CH2 are linked through a flexible hinge region that has the amino-acid sequence APSTCSKPTCP (or APSTCSKPMCP in an allotypic variant) and anchors three disulfide bridges (orange) of the IgG molecule, one for each of the two light- and heavy-chain pairs, and one for the heavy-chain pair. Notably, rabbits have two κ light chains, K1 and K2. The more frequent κ light chain, K1, contains an additional disulfide bridge that links VL and CL. Rabbits of the commonly used New Zealand White strain have ~90% IgG-κ (K1), ~10% IgG-κ (K2) and <1% IgG-λ antibodies. CDR, complementarity-determining region.
Mentions: European rabbits (Oryctolagus cuniculus; Figure 1) have played an important role as animal models in immunology for many decades.1, 2 Today, rabbits are still a major source for a wide variety of monoclonal antibodies (mAbs) and polyclonal antibodies (pAbs) with broad utility. pAbs can be described as a set of different antibodies generated in response to a specific pathogen or antigen, generally targeting different epitopes. mAbs, on the other hand, contain a defined antigen-binding site (paratope) that typically binds with high affinity and specificity to only one epitope. From a pharmaceutical point of view, mAbs provide a molecularly defined and reproducible product, whereas pAbs are traditionally an imprecise mixture of different antibodies.3 As is the case for mouse and human mAbs, IgG is the most common isotype of rabbit mAbs (Figure 2).

View Article: PubMed Central - PubMed

ABSTRACT

In this review, we explain why and how rabbit monoclonal antibodies have become outstanding reagents for laboratory research and increasingly for diagnostic and therapeutic applications. Starting with the unique ontogeny of rabbit B cells that affords highly distinctive antibody repertoires rich in in vivo pruned binders of high diversity, affinity and specificity, we describe the generation of rabbit monoclonal antibodies by hybridoma technology, phage display and alternative methods, along with an account of successful humanization strategies.

No MeSH data available.


Related in: MedlinePlus