GB2103360A - Cell grouping reagent and process - Google Patents

Cell grouping reagent and process Download PDF

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GB2103360A
GB2103360A GB08124244A GB8124244A GB2103360A GB 2103360 A GB2103360 A GB 2103360A GB 08124244 A GB08124244 A GB 08124244A GB 8124244 A GB8124244 A GB 8124244A GB 2103360 A GB2103360 A GB 2103360A
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reagent
saline
protein
cells
agglutinating
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Ian John Deary
Stephen Moore
Duncan Stephen Pepper
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SCOTTISH HEALTH SERVICES ADMIN
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SCOTTISH HEALTH SERVICES ADMIN
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    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/80Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood groups or blood types or red blood cells

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Abstract

A process for the grouping of cells, especially blood cells, consists of adding to a suspension of cells in sodium chloride solution, a cell grouping reagent which has as its active component a saline agglutinating (sa) reagent comprising a non-saline agglutinating (nsa) reagent bound to Protein A, a membrane component of some strains of Staphylococcus aureus. The nsa reagent which binds to Protein A may be for example human or animal monoclonal antibodies or antibodies produced in vitro by human cells, or preferably, human or animal blood cell antibodies, especially IgG. A cell grouping reagent kit comprises an nsa reagent and Protein A, the nsa reagent being either bound to or separate from Protein A. In the latter case, the two components must be mixed (to form an sa reagent) prior to use.

Description

SPECIFICATION Process for the grouping of cells The present invention relates to novel process for the grouping of Cells.
In the process of cell, especially blood cell, grouping, saline reacting ("complete") antisera (those containing saline agglutinating reagents, usually lg M antibodies) have several advantages over the non-saline reacting ("incomplete") antisera (those containing non-saline agglutinating reagents, usually lg G antibodies) which make up the great majority of the non-ABO reagents.
A saline reacting antiserum as defined herein is a serum containing one or more saline agglutinating reagents that react with their opposing cells, causing agglutination, when said cells are suspended in NaCI solution in the absence of antiglobulin serum or enzymes.
By contrast, a non-saline reacting antiserum as defined herein is a serum containing one or more non-saline agglutinating reagents that merely coat their opposing cells, without causing agglutination, when said cells are suspended in NaCI solution in the absence of antiglobulin serum or enzymes. In this latter case the addition of the appropriate antiglobulin serum or enzyme to the coated cells causes the cells to agglutinate.
It can be seen that a saline reacting antiserum and the agglutinating reagents contained therein offer a number of advantages over their non-saline reacting and agglutinating counter parts. For example, a saline reacting antiserum offers a convenient cell especially blood cell grouping test, requiring neither enzyme addition nor an indirect antiglobulin test. Further, the saline reacting antiserum is important in the red cell typing of antibody coated, direct antiglobulin test positive patients.
The saline agglutinating reagents required for saline reacting antisera are seldom found in donors in sufficient quantity or tire for routine use, whilst the purchase of large volumes of commercial reagents or antisera is prohibitively expensive.
There is therefore a need in general for a process for the grouping of cells, in particular blood cells, which, although performed in saline solution in the absence of antiglobulin serum or enzymes, employs the readily available, naturally-occurring non-saline agglutinating reagents rather than the scarce naturally-occurring saline agglutinating reagents.
It is the aim of the present invention to provide a convenient cell grouping test in particular a blood cell grouping test, which may be performed in saline solution without antiglobulin serum or ensymes but which employs naturally-occurring non-saline agglutinating reagents.
According to the present invention there is provided a process for the grouping of cells which comprises adding to a suspension of cells in sodium chloride solution, a cell grouping reagent which has as its active component a saline agglutinating reagent comprising a non-saline agglutinating reagent bound to Protein A (as hereinafter defined).
Preferably the cells to be grouped are blood cells.
The invention further provides a cell grouping reagent for use in the process of the present invention which has as its active component a saline agglutinating (sa) reagent comprising a non-saline agglutinating (nsa) reagent bound to Protein A.
Protein A is a membrane component of some strains of Staphylococcus aureus. It is known to bind specifically to the Fc region of human lg G subclasses 1,2 and 4 while leaving the antigen combining site free to react with the antigen (A Forsgren eft at J Immunol, 1966,97,822; G Kronvall eft at Immunochemistry, 1970, 124).
The Protein A used to bind the nsa reagent may be in substantially pure form or in admixture with, for example, other biologically effective molecules. Preferably however the nsa reagent is bound to Protein A contained in the whole cells of Protein A containing Staphylococcus aureus.
Any nsa reagent which binds to Protein A may be used in the process and cell grouping reagent of the present invention. For example animal or human monoclonal antibodies or antibodies produced in vitro by human cell may be used. Preferably, however, human or animal blood cell antibodies, especially lg G antibodies, are employed. In a particularly preferred embodiment of the process and reagent of the present invention the nsa reagent is a blood cell Ig G antibody specific for antigens in the M-N-S, Rh-Hr, P, Keel, Duffy or kidd blood group systems, for example blood cell lg G, anti-Rho, anti-hr', anti-K, anti-Fya or anti-Jka.
The nsa reagent of the present invention may be in admixture with, for example, other biologically effective compounds. Preferably, however, the nsa reagent will be in substantially pure form. The nsa reagent may be purified by any suitable method, for example by the affinity purification procedure of Rekvig and Hannestad (VoxSang, 1977,33,280).
In the cell grouping reagent of the present invention the sa reagent may be stored in a freeze-dried state, on a solid support, or in solution, for example in phosphate buffered saline (PBS), with or without albumin, or in serum. Generally the sa reagents are stored at temperatures below about 5"C for periods of up to 1-2 weeks without adverse effect on their reactivity.
Once the cell grouping reagent of the present invention has been added to the cell suspension in sodium chloride solution the cell grouping test of the present invention is generally continued by observing whether or not agglutination of the cells takes place.
Generally, if the cell grouping reagent contains, in combination with Protein A, an nsa reagent specific for the antigens of the cells under test, then agglutination will take place. Such agglutination is a visible reaction and therefore may be followed visually in a qualitative or quantitative manner.
On the other hand, if the nsa reagent in the cell grouping reagent is not specific for the cell's antigens then, generally, unless a cross reaction takes place, no agglutination will occur.
Such visual antigen antibody tests are well known in the cell grouping art, forming the basis, in particular, of all blood cell grouping.
In another aspect of the present invention, there are provided certain novel saline agglutinating reagents each comprising a human blood cell (hbc) antibody bound to Protein A contained in the whole cells of Protein A containing Staphylococcus aureus.
Any hbc antibody which binds to Protein A may be used in the novel saline agglutinating reagents of the present invention. For example human blood cell monoclonal antibodies or antibodies produced in vitro by human blood cell. lines may be used. Preferably, however, a human blood cell Ig G antibody, especially those specific for an antigen in the M-N-S, Rh-Hr, P, Kell Duffy or Kidd blood group systems are employed.
Particularly prefered antibodies include human lg G anti-Rho, anti-hr', anti-K, anti-Fya or anti-Jka.
In order to facilitate the routine saline grouping of cells, especially blood cells, it is advantageous for the cell grouping reagents of the present invention to be provided in convenient form for use in medical or blood grouping laboratories. According to a further aspect of the invention therefore there is provided a reagent kit for use in the grouping of cells by the process of the present invention which comprises a saline agglutinating reagent comprising a non-saline agglutinating reagent bound to Protein A.
Alternatively, the reagent kit comprises a. Protein A, and b. a non-saline agglutinating reagent.
In the case of this alternative kit, the Protein A and the nsa reagent are mixed, to form the required sa reagent(nsa reagent bound to Protein A), before use in the cell grouping process of the present invention.
The Protein A used in the reagent kit of the present invention may be in substantially pure form or in admixture with, for example, other biologically effective molecules. Preferably however the Protein A is contained in the whole cells of Protein A containing Staphylococcus aureus.
Any nsa reagent which binds to Protein A may be used in the reagent kit of the present invention. For example animal or human monoclonal antibodies or antibodies produced in vitro by human cell lines may be used. Preferably however human or animal blood cell antibodies, especially lg G antibodies, are employed. Particularly preferred nsa reagents include blood cell lg G antibodies specific for antigens in the M-N-S, Rh-Hr, P, Kell Duffy or Kidd blood group systems, for example blood cell lg G anti-Rho, anti-hr', anti-K, anti-Fya ar anti-Jka.
Preferably the nsa reagent and/or the saline agglutinating reagent of the present reagent kit are in substantially pure form.
Protein A, the sa and/or the nsa reagent may be in freeze-dried form or retained on a solid support. In the preferred reagent kit of the present invention, however, they are dissolved or suspended in one or more suitable solvents. For example, the sa and nsa reagent may be dissolved/suspended in serum or in buffer solution In one particularly preferred embodiment the sa or nsa reagents are dissolved suspended in PBS with albumin added. Similarly, Protein A may be dissolved/suspended in serum or in buffer solution. In one particularly preferred embodiment, the Protein A, is in the form of Pansorbin (Trade Mark), a heat treated, formalinised preparation of Protein A containing S aureus whole cells in 10% suspension (supplied by Calbiochem Behring Corp).
The process and reagents of the present invention will now be described by way of example only.
Materials Pansorbin (TM), a heat treated, formalinised preparation of Staphylococcus aureus in 10% suspension was abtained from Calbiochem, Behring Corp. Membrane ultrafilters were obtained from Amicon Corp. The antibodies used were antisera prepared from human blood donations.
Purification ofnon-saline reacting antibodies Purified antibody was prepared by the adsorption - elution technique of Rekvig and Hannestad, Vox Sang, 1977,33,280. Thus, homozygous cells were washed fourtimes in saline and once in LISS (Low lonic Strength Saline, NaCI, 7.1 6g; NaH2 P04, 1 .87g; glycine, 729; all dissolved to 4 litre in distilled water; pH 6.7 with NaOH aq). Packed cells were resuspended to 50% in LISS and 1 vol of antibody was added. The mixture was incubated at 37"Cfor 1 hr and the cells were then washed four times in a large excess of phosphate buffered saline (PBS, pH 7.4) and finally resuspended to 50% in the same buffer. Antibody was eiuted from the sensitised cells, previously cooled to OOC, by rapid addition of 1 vol of ice-cold elution buffer (0.1 M glycine, Q.1 M NaCI, pH adjusted to 2.8 with H CI) followed by thorough mixing and immediate centrifugation (32,Q00 xg) at 4"C for 30 seconds. The supernatant was collected and the pH immediately adjusted to pH 7.5 with a concentrated tris solution. A second eiution was performed immediately. Both eluates were pooled and concentrated by ultrafiltration on an Amicon XM 50 membrane.
Antibody quantitation Antibody quantitation was performed by using a modified Marsh auto-analyser procedure (Gunson, Phillips and Stratton, dC/in Path, 1972,25, 198).
Agglutination test Agglutinating ability was tested by adding 1 vol of a 5% cell suspension in 0.9% NaCI to 1 vol of saline agglutinating reagent (Protein A + nsa reagent). To rule out non-specific agglutination due to the saline agglutinating reagent, negative controls were set up in parallel with all tests, these were as indicated above except that the cells lacked the antigen against which the saline agglutinating reagent was directed. Tests were performed in 3 inch by 3/8 inch glass tubes, incubated at 37"C for 45 mins, centrifuged at 500 xg for 3 sec and read macroscopically.Agglutination strength was scored using the following convention: Complete agglutination = 4 +++ = 3 ++ = 2 + = 1 (+) = 0.5 (+) = 0.25 Production of antibody - sensitised, direct antiglobulin test positive cells This was performed by adding 2 vols of antiserum to 1 vol of an appropriate 5% cell suspension in 0.9% NaCI. After incubating for 1 hr at 37 C the cells were washed four times in 0.9% NaCI and finally made up to a 5% suspension in 0.9% NaCI, readyfortesting.
EXAMPLE 1 Preparation ofa saline agglutinating reagent by reacting a nsa reagent with whole cells of protein A containing Staphylococcus aureus Anti-Rho, in the form of a concentrated eluate, was produced by the purification (of nsa reagents) procedure described above. The starting volume of "incomplete" antibody was 25 ml and the % eluate recovery (calculated from the anti-Rho quantitation of starting material and eluate) was 71%.
Pansorbin (Trade Mark, 0.25 ml) was then added to the concentrated eluate containing anti-Rho, and the mixture was incubated with occasional mixing at 370 for 1 hr. The mixture was then centifuged and the S aureus cells were washed, once in a solution of 1% Triton X - 100 2M NaCI, 0.01 M phosphate, pH 7.4, to remove non-specifically bound protein, and twice in PBS pH 7.4 before being resuspended in PBS pH 7.4 + 1% albumin to a volume of 0.5 ml.
The activity of the starting material (concentrated eluate of anti-Rho) and of the anti-Rho bound to Protein A (PA, anti-Rho) versus R2 R2 cells in saline medium, using the agglutination test described above, is shown in Table I.
EXAMPLE 2 The procedure of Example 1 was carried out exceptforthefollowing modifications.
i. The starting volume of "incomplete" antibody was 12 ml.
ii. The % eluate recovery was 44% iii. The volume of Pansorbin added was 0.1 ml iv. The (PA, anti-Rho) was resuspended in PBS pH 7.4 + 1% albumin to a volume of 0.4 ml.
The activity of the starting material and of the (PA, anti-Rho) versus R2R2 cells in saline medium, using the agglutination test described above, is shown in Table I.
EXAMPLE 3 The procedure of Example 1 was carried out except for the following modifications.
i. The starting volume of "incomplete" antibody was 12 ml.
ii. The % eluate recovery was 42%.
iii. The (PA, anti-Rho) was resuspended in PBS pH 7.4 + 1% albumin to a volume of 1.0 ml.
The activity of the starting material and of the (PA, anti-Rho) versus R2R2 cells in saline medium, using the agglutination test described above, is shown in Table I.
EXAMPLE 4 The procedure of Example 1 was carried out except for the following modification.
i. The starting volume of "incomplete" antibody was 22 ml.
ii. The % eluate recovery was 47%.
The activity of the starting material and of the (PA, anti-Rho) versus R2 R2 cells in saline medium, using the agglutination test described above, is shown in Table I.
EXAMPLE 5 The procedure of Example 1 was carried out except for the following modifications.
i. The starting volume of "incomplete" antibody was 60 ml.
ii. The volume of Pansorbin added was 0.5 ml.
iii. The (PA, anti-Rho) was resuspended in PBS pH 7.4 + 1% albumin to a final volume of 1.0 ml.
The activity of the starting material and of the (PA, anti-Rho) versus R0 r cells in saline medium, using the agglutination test described above, is shown in Table I.
EXAMPLE 6 The procedure of Example 1 was carried out except for the following modifications.
i. Anti-K replaced anti-Rho.
ii. The starting volume of "incomplete" antibody was 50 ml.
The activity of the starting material and of the (PA, anti-K) versus Kk cells in saline medium, using the agglutination test described above, is shown in Table I.
EXAMPLE 7 The procedure of Example 1 was carried out except for the following modifications.
i. Anti-hr' replaced anti-Rho.
ii. The starting volume of "incomplete" antibody was 50 ml.
The activity of the starting material and of the (PA, anti-hr') versus R2 R2 cells in saline medium, using the agglutination test described above, is shown in Table I.
TABLE I Example Starting material activitya (PA, antibody) activitya Saline Titre ICTb Saline Titre ICTb 1 None 512 64 128 (20.5)C (12) (17) 2 None 512 8 64 (20.5) (4.5) (14.5) 3 None 512 16 64 (20.5) (5.5) (11.5) 4 None 512 64 64 (20.5) (11.5) (13.5) 5 None 256 16 16 (20.5) (3.75) (4.5) 6 None 128 16 64 (17) (4) (5) 7 None 128 16 16 (20.5) (5) (5) Notes: a, Results are reciprocal titration values b, Indirect antiglobulinetest performed on saline tests.
c, Figures in parentheses are the cumulative agglutination scores.
EXAMPLE 8 The procedure of Example 1 was carried out except for the following modifications.
The starting volume of "incomplete" antibody was 100 ml.
ii. The volume of Pansorbin added was 1 ml.
iii. The (PA, anti-Rho) was resuspended in PBS pH 7.4 + 1% albumin to a final volume of 2.0 ml.
The resuspended (PA, anti-Rho) was then split into three samples, A, B and C.
The activity of sample A versus R2 R2 cells was ascertained immediately after the (PA, anti-Rho) was prepared.
The activity of sample B versus R2 R2 cells was ascertained after storage at 40C for 1 week.
The activity of sample C versus R2 R2 cells was ascertained after the sample had been freeze-dried, stored at -20 C for 1 week and then reconstituted.
The activities of samples A, B and C are shown in Table II.
TABLE II Sample Saline Titrea ICTb A 128 512 (15.75)C (20.5) B 128 512 (10.5) (22) C 128 512 (23.5) (26.5) Note: a, Results are reciprocal titration values b, Indirect antiglobulin test performed on saline tests c, Figures in parantheses are the cumulative agglutination scores.
EXAMPLE 9 The procedure of Example 7 was carried out. The resuspended (PA, anti-hr') was then split into two samples D and E.
The activity of sample D versus R2 R2 cells was ascertained immediately after the (PA, anti-hr') was prepared.
The activity of sample E versus R2 R2 cells was ascertained after storage at 40C for 1 week.
The activities of samples D and E are shown in Table Ill.
TABLE III Sample Saline Titrea D 16 (5)C E 16 (5.25) a, Results are reciprocal titration values, c, Figures in parentheses are the cumulative agglutination scores.
EXAMPLE 10 The procedure of Example 1 was carried out to prepare (PA, anti-Rho) suspended in PBS pH 7.4 + 1% albumin atatitre of 1/6.
This reagent was then used to Rho type cells which were antibody coated, direct antiglobulin test positive.
(The cells were produced as described above under the production of antibody-sensitised, direct antiglobulin test positive cells). The cells under test were Rho positive and Rho negative cells sensitised with anti-rh" anti-Fy0 or anti-K ell.
The results of these agglutination tests are given in Table IV.
TABLE IV The use of (PA, anti-Rho) in Rho typing of antibody - sensitised, direct antiglobullin test positive cells.
Cell Sensitising Antibody Reaction with Reaction with AHGa (PA, anti-Rho) R2 R2 anti-rh" +++ ++ R"r anti-rh" Rho+, Fya anti-Fy0 + ++ Rho-, Fya anti-Fy0 (+) Rho+, Kk anti-Kell + Rho-, Kk anti-Kell + Note: a, anti-human globulin reagent CLAIMS (Filed on 23 July 1982) 1. A process for the grouping of cells comprises adding to a suspension of cells in sodium chloride solution, a cell grouping reagent which has as its active component a saline agglutinating reagent comprising a non-saline agglutinating reagent bound to protein A (as hereinbefore defined).
2. A process according to claim 1 wherein the cells to be grouped are blood cells.
3. A process according to either claim 1 or claim 2 wherein the saline agglutinating reagent comprises a non-saline agglutinating reagent bound to Protein A contained in the whole cells of Protein A containing Staphylococcus aureus.
4. A process according to any one of claims 1 to 3 wherein the non-saline agglutinating reagent comprises human or animal blood cell antibodies.
5. A process according to claim 4 wherein the non-saline agglutinating reagent comprises IgG antibodies.
6. A process according to claim 5 wherein the non-saline agglutinating reagent comprises blood cell IgG antibodies specific for antigens in the M-N-S, Rh - Hr, P, Kell, Duffy or Kidd blood group systems.
7. A process according to claim 6 wherein the blood cell IgG antibodies are selected from anti - Rho, anti hr', anti - K, anti - Fya and anti - Jka.
8. A process for the grouping of cells according to claim 1 substantially as hereinbefore described with particular reference to any one of the Examples.
9. A cell grouping reagent comprising an active component in conjunction with an acceptable diluent or carrier wherein the active component is a saline agglutinating reagent comprising a non-saline agglutinating reagent bound to Protein A (as hereinbefore defined).
10. A cell grouping reagent according to claim 9 substantially as hereinbefore described with particular reference to any one of the Examples.
11. A cell grouping reagent kit for use in the grouping cells by a process according to claim 1 comprising a cell grouping reagent according to either claim 9 or claim 10.
12. A cell grouping reagent kit for use in the grouping of cells by a process according to claim 1 comprising Protein A (as hereinbefore defined) and a non-saline agglutinating reagent.
13. A cell grouping reagent kit according to either claim 11 or claim 12 wherein the Protein A is contained in the whole cells of Protein A containing Staphylococcus aureus.
14. A cell grouping reagent kit according to claim 13 wherein the Protein A is in the form of a heat-treated, formalinised preparation of Protein A containing Staphylococcus aureus whole cells in 10% suspension.
15. A cell grouping reagent kit according to any one of claims 11 to 14 wherein the non-saline agglutinating reagent comprises human or animal blood cell antibodies.
16. A cell grouping reagent kit according to claim 15 wherein the non-saline agglutinating reagent comprises IgG antibodies.
17. A cell grouping reagent kit according to claim 16 wherein the non-saline agglutinating reagent comprises blood cell IgG antibodies specific for antigens in the M-N-S, Rh - Hr, P, kell, duffy or Kidd blood group systems.
18. A cell grouping reagent kit according to claim 17 wherein the blood cell IgG antibodies are selected from anti-Rho, anti-hr', anti - K, anti - Fya and anti - Jka.
19. A cell grouping reagent kit according to any one of claims 11 to 18 wherein the saline agglutinating reagent or the non-saline agglutinating reagent is, in each case, either dissolved or suspended in phosphate buffered saline with albumin added.
20. A cell grouping reagent kit according to either claim 11 or 12 substantially as hereinbefore described with particular reference to any one of the Examples.
21. A saline agglutinating reagent comprising a human blood cell antibody bound to Protein A (as hereinbefore defined) contained in the whole cells of Protein A containing Staphylococcus aureus.
22. A saline agglutinating reagent according to claim 21 substantially as herein before described with particular reference to any one of the Examples.
New claims or amendments to claims filed on 8 September 1982 Superseded claims 1 to 22 New or amended claims:

Claims (22)

1. A process for the grouping of cells comprises adding to a suspension of cells in sodium chloride solution, a cell grouping reagent which has as its active component a saline agglutinating reagent comprising a non-saline agglutinating reagent bound to protein A (as hereinbefore defined).
2. A process according to claim 1 wherein the cells to be grouped are blood cells.
3. A process according to either claim 1 or claim 2 wherein the saline agglutinating reagent comprises a non-saline agglutinating reagent bound to Protein A contained in the whole cells of Protein A containing Staphylococcus aureus.
4. A process according to any one of claims 1 to 3 wherein the non-saline agglutinating reagent comprises human or animal blood cell antibodies.
5. A process according to claim 4 wherein the non-saline agglutinating reagent comprises IgG antibodies.
6. A process according to claim 5 wherein the non-saline agglutinating reagent comprises blood cell IgG antibodies specific for antigens in the M-N-S, Rh, - Hr, P, Kell, Duffy or Kidd blood group systems.
7. A process according to claim 6 wherein the blood cell IgG antibodies are selected from anti - Rho, anti hr', anti - K, anti - Fya and anti - Jka.
8. A process for the grouping of cells according to claim 1 substantially as hereinbefore described with particular reference to any one of the Examples.
9. A cell grouping reagent comprising an active component in conjunction with an acceptable diluent or carrier wherein the active component is a saline agglutinating reagent comprising a non-saline agglutinating reagent bound to Protein A (as hereinbefore defined).
10. A cell grouping reagent according to claim 9 substantially as hereinbefore described with particular reference to any one of the Examples.
11. A cell grouping reagent kit for use in the grouping of cells by a process according to claim 1 comprising a cell grouping reagent according to either claim 9 or claim 10.
12. A cell grouping reagent kit for use in the grouping of cells by a process according to claim 1 comprising Protein A (as hereinbefore defined) and a non-saline agglutinating reagent.
13. A cell grouping reagent kit according to either claim 11 or claim 12 wherein the Protein A is contained in the whole cells of Protein A containig Staphylococcus aureus.
14. A cell grouping reagent kit according to claim 13 wherein the Protein A is in the form of a heat-treated, formalinised preparation of Protein A containing Staphylococcus aureus whole cells in 10% suspension.
15. A cell grouping reagent kit according to any one of claims 11 to 14 wherein the non-saline agglutinating reagent comprises human or animal blood cell antibodies.
16. A cell grouping reagent kit according to claim 15 wherein the non-saline agglutinating reagent comprises IgG antibodies.
17. A cell grouping reagent kit according to claim 16 wherein the non-saline agglutinating reagent comprises blood cell IgG antibodies specific for antigens in the M-N-S, Rh - Hr, P, kell, Duffy or Kidd blood group systems.
18. A cell grouping reagent kit according to claim 17 wherein the blood cell IgG antibodies are selected from anti-Rho, anti-hr', anti - K, anti - Fya and anti - Jka.
19. A cell grouping reagent kit according to any one of claims 11 to 18 wherein the saline agglutinating reagent or the non-saline agglutinating reagent is, in each case, either dissolved or suspended in phosphate buffered saline with albumin added.
20. A cell grouping reagent kit according to either claim 11 or 12 substantially as hereinbefore described with particular reference to any one of the Examples.
21. Asaline agglutinating reagent comprising a human blood cell antibody bound to Protein A (as hereinbefore defined) contained in the whole cells of Protein A containing Staphylococcus aureus.
22. A saline agglutinating reagent according to claim 21 substantially as herein before described with particular reference to any one of the Examples.
GB08124244A 1981-08-07 1981-08-07 Cell grouping reagent and process Withdrawn GB2103360A (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2676123A1 (en) * 1991-05-02 1992-11-06 Pasteur Diagnostics AGGLUTINANT COMPLEX AND REAGENT FOR IDENTIFYING ANTIGENS ON CELL WALLS.
CN112469109A (en) * 2020-12-02 2021-03-09 Oppo广东移动通信有限公司 Method for preferentially selecting ENDC cell, terminal device and storage medium

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2676123A1 (en) * 1991-05-02 1992-11-06 Pasteur Diagnostics AGGLUTINANT COMPLEX AND REAGENT FOR IDENTIFYING ANTIGENS ON CELL WALLS.
EP0512896A1 (en) * 1991-05-02 1992-11-11 Pasteur Sanofi Diagnostics Agglutination complex for the determination of blood groups
CN112469109A (en) * 2020-12-02 2021-03-09 Oppo广东移动通信有限公司 Method for preferentially selecting ENDC cell, terminal device and storage medium

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