Construction and preliminary identification of a prokaryotic expression single-chain antibody fragments library against Streptococcus pneumoniae from antibody-producing cells in human tonsil

Figures & data

Table 1. scfv sequence alignment on ClustaIW.

SeqA	Length	SeqB	Length	Score
1	775	2	776	72.72
1	775	3	761	77
1	775	4	752	68.75
1	775	5	736	67.53
1	775	6	773	72.19
1	775	7	757	82.96
1	775	8	793	81.68
1	775	9	773	72.19
1	775	10	792	88.26
2	776	3	761	68.2
2	776	4	752	69.68
2	776	5	736	73.37
2	776	6	773	72.72
2	776	7	757	79.66
2	776	8	793	80.42
2	776	9	773	72.72
2	776	10	792	74.15
3	761	4	752	75.13
3	761	5	736	70.38
3	761	6	773	67.15
3	761	7	757	68.82
3	761	8	793	66.89
3	761	9	773	67.15
3	761	10	792	75.69
4	752	5	736	78.4
4	752	6	773	77.79
4	752	7	757	68.48
4	752	8	793	69.02
4	752	9	773	77.79
4	752	10	792	68.62
5	736	6	773	63.72
5	736	7	757	72.96
5	736	8	793	77.17
5	736	9	773	63.72
5	736	10	792	68.89
6	773	7	757	75.69
6	773	8	793	72.7
6	773	9	773	100
6	773	10	792	70.63
7	757	8	793	88.51
7	757	9	773	75.69
7	757	10	792	82.96
8	793	9	773	72.7
8	793	10	792	83.08
9	773	10	792	70.63

Table 2. The results of scfv sequence analysis on IGBLAST.

Scfv sequences	Top matched V gene
	VH VL
1	IGHV1-69*09 (94.9%)	IGLV2-23*02(97.6%)
2	IGHV3-30–3*01 (91.2%)	IGLV6-57*02 (100.0%)
3	IGHV1-69*14 (93.2%)	IGKV3-15*01 (91.9%)
4	IGHV4-39*07 (96.0%)	IGKV1-39*01 (98.2%)
5	IGHV3-7*01 (92.5%)	IGKV1D-33*01 (96.0%)
6	IGHV4-38–2*02 (96.9%)	IGLV3-21*03 (94.8%)
7	IGHV3-66*01 (95.9%)	IGLV2-14*01 (93.1%)
8	IGHV3-30*18 (91.2%)	IGLV2-14*01 (96.8%)
9	IGHV4-38–2*02 (96.9%)	IGLV3-21*03 (94.8%)
10	IGHV1-2*02 (97.6%)	IGLV2-14*01 (94.9%)

Figure 1. Map of pMo Pac16 vector.

Figure 2. The result of an indirect ELISA assay for detection of the IgG against S. aureus, group A Streptococcus, beta Streptococcus, and S. pneumoniae (n = 3).

Figure 3. (a) Amplification of VH linker, Vλ linker, and Vκ linker gene by PCR, M: DNA marker DL2000. (1) PCR product of VH linker. (2) PCR product of Vκ linker. (3) PCR product of Vλ linker. (b) Purified PCR products of scfv fragment. M, DNA marker DL2000 1: purified PCR products of scfv. (c) The identification of recombinant clones by SfiI. M: DNA marker DL15000. Channel 1–8, the identification of pMo Pac16-scFv by Sfi.

Figure 4. (a) ELISA assay of binding activity for scfv. (1) Blank control; (2) negative control (null vector transformant), 3 ~ 69 scfv. (b) ELISA assay of specificity for positive scfv (n = 3).