Allotype related charge differences between immunoglobulins and light chains from rabbits differing at the Ab locus

Notenboom, Robert; Dubiski, S.; Cinader, B.; Underdown, Brian J.

doi:10.1016/0161-5890(79)90047-6

Cited by 4 publications

(3 citation statements)

References 25 publications

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“…Elution volume ( Characterization of the hybridornu protein The PI value of the purified hybridoma protein ( 5 mg) in the absence of urea was 6.1, a value within the range of IgG of allotype Ab4 as previously determined (Notenboom et al, 1979).…”

Section: Purification Of Hybridoma Proteinsupporting

confidence: 62%

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Isolation and Characterization of a Mouse‐rabbit Hybridoma

Notenboom

Chou

Good

et al. 1980

Int J Immunogenetics

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Summary Immunoglobulin was obtained from a hybridoma cell line, which was a reclone of a hybrid between a rabbit cell and mouse myeloma cell (X63‐Ag8). The immunoglobulin, isolated from the cell culture medium was found to be homogeneous by isoelectrofocusing and immunoelectrophoresis and consisted of mouse heavy and rabbit light chains, linked by disulphide bonds. All immunoglobulin molecules carried both mouse and rabbit determinants; mouse determinants were associated only with the heavy chains while rabbit determinants were only associated with the light chains. The rabbit light chains were of Ab4 allotypical specificity, but possess only some of the Ab4 determinants normally present in Ab4/Ab4 animals. It was sugested that the restriction in allotypical specificity may be a general property of light chain Ab allotypes; the normal serum immunoglobulins may be heterogeneous with respect to the allotypic determinants and any one molecule may possess only a proportion of determinants detected by a conventional anti‐allotype antiserum.

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Section: Purification Of Hybridoma Proteinsupporting

confidence: 62%

“…(rnmunoeiectrop horesis used. f soeiectrofoczrsing has been described previously (Notenboom et al, 1979).…”

Section: Gel Chroinatographymentioning

confidence: 99%

Isolation and Characterization of a Mouse‐rabbit Hybridoma

Notenboom

Chou

Good

et al. 1980

Int J Immunogenetics

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“…The initial observation, relating antigen charge to antibody charge (Sela and Mozes, 1966), was made with rabbits, presumably homozygous with respect to the predominant light chain allotype, Ab4. In our study of charge differences between rabbit light chains of various allotypes, we found that the charge spectrum of Ab4 was much wider than that of Ab9 (Notenboom et al, 1979). This led us to the view that the structural genes of the light chain might determine the probability of effective contact between Ig receptor and antigen (Szymanska et al, 1981) In rabbits heterozygous at the kappa light chain allotypic locus (Ab) the two allelic genes are not equally expressed.…”

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confidence: 99%

Allotype and Charge‐related Differences in the Immune Response

Notenboom

Dubiski

and

et al. 1982

Int J Immunogenetics

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SUMMARY Rabbits homozygous at the Ab kappa chain allotypic locus (Ab4/Ab4 and Ab9/Ab9) were immunized with negatively and positively charged antigens. The negatively charged antigens were bovine serum albumin (BSA; pI = 4.9), ovalbumin (OV; pI = 4.9) and two synthetic polypeptides: copolymer L‐glutamic acid L‐tyrosine (abbreviated TG; pI = 4.8) and multichain poly (L‐tyrosine: L‐glutamic acid) poly D‐alanine: poly L‐lysine (poly L‐lysine backbone) (abbreviated (TG) ‐ AL; pI = 4.8). The positively charged antigen was hen egg lysozyme (LYS; pI = 10.2). Homozygous Ab9 rabbits responding to negatively charged antigens made less antibody than did Ab4 homozygotes. In contrast, both groups of animals responded equally well to positively charged lysozyme. The relative electric charges of the antibodies were assessed by Sephadex A‐50 chromatography. The charge distribution was found to depend on the charge of the eliciting antigen. The positively charged antibodies of Ab9/Ab9 rabbits were not nearly as positively charged as those raised in Ab4/Ab4 rabbits. The difference in average electric charges and in the range of these charges between Ig Ab4 and Ig Ab9 explain quantitative differences in antibody formation in response to negatively charged antigens and may be a contributing factor to the ‘pecking order’, i.e. unequal phenotypic expression of ligh chain genes in Ab4/Ab9 heterozygotes.

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